Tomato variety nun 00316 top

ABSTRACT

A new and distinct tomato variety NUN 00316 TOP is disclosed, as well as seeds and plants and fruits thereof. NUN 00316 TOP is a determinate tomato variety of the viscosity type for the processing market, comprising resistance to  Verticillium dahliae, Fusarium oxysporum  s.p.  lycopersici  Race 2 (ex 3),  Meloidogyne incognita, Pseudomonas syringae  pv tomato,  Leveillula taurica,  and Tomato Spotted Wilt Virus (TSWV).

CROSS-REFERENCE TO RELATED APPLICATION

This application claims priority of U.S. Provisional Application No.63/126,930 filed on Dec. 17, 2020, which is hereby incorporated byreference in its entirety.

FIELD OF DISCLOSURE

The disclosure relates to the field of plant breeding, more specificallyto tomato variety NUN 00316 TOP. The disclosure further relates tovegetative reproductions of tomato variety NUN 00316 TOP, methods fortissue culture of tomato variety NUN 00316 TOP, methods for regeneratinga plant from such a tissue culture, and to phenotypic variants of tomatovariety NUN 00316 TOP. The disclosure also relates to progeny of tomatovariety NUN 00316 TOP as a parent line with plants of other varieties orparent lines.

BACKGROUND OF THE DISCLOSURE

The goal of plant breeding is to combine various desirable traits in asingle variety. Such desirable traits may include greater yield,resistance to diseases, insects or other pests, tolerance to heat anddrought, better agronomic quality, higher nutritional value, enhancedgrowth rate and improved fruit properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same genotype. A plant cross-pollinates if pollen comes to it from aflower of a different genotype.

Plants that have been self-pollinated and selected for (uniform) typeover many generations become homozygous at almost all gene loci andproduce a uniform population of true breeding progeny of homozygousplants. A cross between two such homozygous plants of different linesproduces a uniform population of hybrid plants that are heterozygous formany gene loci. The extent of heterozygosity in the hybrid is a functionof the genetic distance between the parents. Conversely, a cross of twoplants each heterozygous at a number of loci produces a segregatingpopulation of hybrid plants that differ genetically and are not uniform.The resulting non-uniformity makes performance unpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants to makehybrids, and the evaluation of the hybrids resulting from the crosses.Pedigree breeding and recurrent selection are examples of breedingmethods that have been used to develop inbred plants from breedingpopulations. Those breeding methods combine the genetic backgrounds fromtwo or more plants or various other broad-based sources into breedingpools from which new lines are developed by selfing and selection ofdesired phenotypes. The new plants are evaluated to determine which havecommercial potential.

Tomato (Solanum lycopersicum and closely related species) is naturally adiploid and the basic chromosome number of the genus is x=12, most are2n=2x=24, including the cultivated ones. It originated in the New Worldand has since become a major food crop.

Tomato cultivars may be grouped by maturity, i.e., the time requiredfrom planting the seed to the stage where fruit harvest can occur.Standard maturity classifications include ‘early’ ‘midseason’ or‘late-maturing’. Another classification for tomatoes is thedevelopmental timing of fruit set. ‘Determinate’ plants grow foliage,then transition into a reproductive phase of flower setting, pollinationand fruit development. Consequently, determinant cultivars have a largeproportion of the fruit ripen within a short time frame. Growers thatharvest only once in a season favor determinant type cultivars. Incontrast, ‘indeterminate’ types grow foliage, then enter a long phasewhere flower and fruit development proceed along with new foliar growth.Growers that harvest the same plants multiple times favor indeterminatetype cultivars.

Tomatoes can also be classified by their target markets: fresh marketand processing tomatoes. Fresh-market tomatoes are primarily used forsalads, salad bar, and sandwiches, and require good storage properties.On the other hand, processing tomatoes generally requires red coloredand pink to red/crimson fruit flesh and higher percentage of solublesolids. Processing tomatoes can be canned whole, canned, diced orchopped, dried, roasted, pasted, puréed or concentrated, juiced, frozen,or put into ready-made dishes, for example, sauces, stews, or soups.

In 2017, World Atlas (available at world-wide web word atlas underarticles/which-are-the-world-s-leading-tomato-producing-countries)reported that the worldwide production of tomatoes amounted to 170.8million tons. United States is ranked as the third largest producer oftomatoes in the world, next to China and India. Tomatoes are availablein the United States year-round, with California and Florida being themajor producers. Fresh-market tomatoes are available from May toDecember although supply peaks in July and in September through October.Processing tomatoes have the greatest supply from August to September.

In response to more recent consumer demands for dietary diversity,tomato breeders have developed a wider range of colors. In addition toexpanding the range of red colored fruits, there are cultivars thatproduce fruits that are creamy white, lime green, yellow, green, golden,orange, and purple. Additionally, there are multi-colored varietiesexemplified by mainly red fruited varieties with green shoulders, andboth striped- and variegated-colored fruit.

SUMMARY OF THE VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure provides for tomato variety NUN 00316 TOP, productsthereof, and methods of using the same. NUN 00316 TOP is a determinatetomato variety of the viscosity type for the processing market and issuitable for growing in the open field.

The disclosure also provides a tomato plant or part thereof having allof the physiological and morphological characteristics of tomato varietyNUN 00316 TOP when grown under the same environmental conditions.

In another aspect, the plant of tomato variety NUN 00316 TOP or aprogeny thereof comprises resistance to Verticillium dahliae, Fusariumoxysporum s.p. lycopersici Race 2 (ex 3), Meloidogyne incognita,Pseudomonas syringae pv tomato, Leveillula taurica, and Tomato SpottedWilt Virus (TSWV), measured according to UPOV standards described inTG/44/11.

The disclosure also provides for a progeny of tomato variety NUN 00316TOP. In a further aspect, the plant or progeny retains all or all butone, two, or three of the “distinguishing characteristics” or all or allbut one, two, or three of the “morphological and physiologicalcharacteristics” of tomato variety NUN 00316 TOP and methods forproducing that plant or progeny.

In another aspect, the disclosure provides a plant or a progeny havingall the physiological and morphological characteristics of variety NUN00316 TOP when grown under the same environmental conditions. In anotheraspect, the plant or progeny has all or all but one, two, or three ofthe physiological and morphological characteristics of tomato varietyNUN 00316 TOP when measured under the same environmental conditions ande.g., evaluated at significance levels of 1%, 5%, or 10% significance(which can also be expressed as a p-value) for quantitativecharacteristics and determined by type or degree for non-quantitativecharacteristics, wherein a representative sample of seed of tomatovariety NUN 00316 TOP has been deposited under Accession Number NCIMB______. In another aspect, the plant or progeny has all or all but one,two, or three of the physiological and morphological characteristics aslisted in Tables 1-3 for variety NUN 00316 TOP when measured under thesame environmental conditions and e.g., evaluated at significance levelsof 1%, 5%, or 10% significance (which can also be expressed as ap-value) for quantitative characteristics and identical (same type ordegree) for non-numerical characteristics.

In another aspect, the plant of tomato variety NUN 00316 TOP, or partthereof, has at least 23, 24, or more of the following distinguishingcharacteristics when compared to its Reference Variety as shown in Table4: 1) shallowly toothed or scalloped margin of leaflets; 2) strongermarginal leaf rolling or wiltiness; 3) shorter leaf; 4) narrower leaf;5) smaller petiole; 6) lighter green leaf color; 7) shorter pedicel fromjoint to calyx; 8) larger fruit diameter; 9) heavier fruit; 10) three tofour locules; 11) lighter green fruit base color at mature-green stage;12) lower % soluble solids; 13) lighter intensity of green colorexcluding shoulder before maturity; 14) lighter yellow green color ofgreen fruit; 15) larger fruit size; 16) obovate fruit shape inlongitudinal section; 17) stronger depression at peduncle end; 18) flatto pointed shape of blossom end; 19) larger core in cross-section inrelation to total diameter; 20) darker red color of mature fruit; 21)darker red color of flesh at maturity; 22) larger pedicel; 23) resistantto Leveillula taurica; and 24) resistant to Pseudomonas syringae pvtomato, when the numerical characteristics are determined at 5%significance level and determined by type or degree for non-numericalcharacteristics when grown under the same environmental conditions.

In another aspect, the disclosure provides a seed of tomato variety NUN00316 TOP, wherein a representative sample of said seed has beendeposited under Accession Number NCIMB ______. The disclosure alsoprovides for a plurality of seeds of tomato variety NUN 00316 TOP. Thetomato seed of variety NUN 00316 TOP may be provided as an essentiallyhomogeneous population of tomato seed. The population of seed of tomatovariety NUN 00316 TOP may be particularly defined as being essentiallyfree from other seed. The seed population may be grown into plants toprovide an essentially homogeneous population of tomato plants asdescribed herein.

The disclosure also provides a plant grown from a seed of tomato varietyNUN 00316 TOP and a plant part thereof.

The disclosure also provides a tomato fruit produced on a plant grownfrom a seed of tomato variety NUN 00316 TOP.

The disclosure furthermore provides a seed growing or grown on a plantof variety NUN 00316 TOP (i.e., produced after pollination of the flowerof tomato variety NUN 00316 TOP).

In another aspect, the disclosure provides for a plant part obtainedfrom tomato variety NUN 00316 TOP, wherein said plant part is: a fruit,a harvested fruit, a part of a fruit, a leaf, a part of a leaf, pollen,an ovule, a cell, a petiole, a shoot or a part thereof, a stem or a partthereof, a root or a part thereof, a root tip, a cutting, a seed, a partof a seed, seed coat or another maternal tissue which is part of a seedgrown on said variety, a hypocotyl, a cotyledon, a scion, a stock, arootstock, a pistil, an anther, or a flower or a part thereof. Fruitsare particularly important plant parts. In another aspect, the plantpart obtained from tomato variety NUN 00316 TOP is a cell, optionally acell in a cell or tissue culture. That cell may be grown into a plant ofvariety NUN 00316 TOP.

In another aspect, the disclosure provides a hybrid tomato variety NUN00316 TOP.

In another aspect, the disclosure provides an inbred variety of tomatovariety NUN 00316 TOP.

The disclosure also provides a cell culture of tomato variety NUN 00316TOP and a plant regenerated from tomato variety NUN 00316 TOP, whereinthe plant has all or all but one, two, or three of the characteristicsof tomato variety NUN 00316 TOP when grown under the same environmentalconditions, as well as methods for culturing and regenerating tomatovariety NUN 00316 TOP. Alternatively, a regenerated plant may have onecharacteristic that is different from tomato variety NUN 00316 TOP andotherwise has all the physiological and morphological characteristics oftomato variety NUN 00316 TOP when grown the same environmentalconditions and wherein a representative sample of seed of said tomatovariety has been deposited under Accession Number NCIMB ______.

The disclosure further provides a vegetatively propagated plant ofvariety NUN 00316 TOP having all or all but one, two, or three of themorphological and physiological characteristics of tomato variety NUN00316 TOP when grown under the same environmental conditions as well asmethods for vegetatively propagating tomato variety NUN 00316 TOP.

In another aspect, the disclosure provides a method of producing atomato plant comprising crossing tomato variety NUN 00316 TOP withitself or another tomato variety and selecting a progeny tomato varietyfrom said crossing or selfing.

The disclosure also provides a method of producing a tomato plantderived from tomato variety NUN 00316 TOP.

In a further aspect, the disclosure provides a method of producing ahybrid tomato seed comprising crossing a first parent tomato plant witha second parent tomato plant and harvesting the resultant hybrid tomatoseed, wherein said first parent tomato plant or second parent tomatoplant is tomato variety NUN 00316 TOP. Also provided is a hybrid tomatoseed produced from crossing a first parent tomato plant with a secondparent tomato plant and harvesting the resultant seed, wherein saidfirst parent tomato plant or second parent tomato plant is tomatovariety NUN 00316 TOP. Moreover, the hybrid tomato plant grown from thehybrid tomato seed is provided.

In another aspect, the disclosure provides a method of introducing asingle locus conversion into the plant of variety NUN 00316 TOP, whereina representative sample of seed of said tomato variety has beendeposited under Accession Number NCIMB ______, wherein the plantcomprises the single locus conversion and otherwise has all of themorphological and physiological characteristics of tomato variety NUN00316 TOP.

In yet another aspect, the disclosure provides a method of introducing adesired trait into tomato variety NUN 00316 TOP, said method comprisestransforming the plant of variety NUN 00316 TOP with a transgene thatconfers the desired trait, wherein the transformed plant contains thedesired trait and otherwise has all of the morphological andphysiological characteristics of tomato variety NUN 00316 TOP.

The disclosure also provides a method of producing a modified tomatovariety with a desired trait, wherein the method comprises mutating thetomato plant or plant part of tomato variety NUN 00316 TOP, wherein arepresentative sample of seed of said tomato variety has been depositedunder Accession Number NCIMB ______, and wherein the mutated plantcontains the desired trait and otherwise has all of the morphologicaland physiological characteristic of tomato variety NUN 00316 TOP.

In one aspect, the single locus conversion or desired trait is yield,size, shape, color, flavor or taste, storage properties, nutritionalquality, post-harvest quality, male sterility, herbicide tolerance,insect resistance, pest resistance, disease resistance, environmentalstress tolerance, modified carbohydrate metabolism, modified proteinmetabolism, or ripening.

In another aspect, the disclosure provides a container comprising theplant, plant part, or seed of tomato variety NUN 00316 TOP.

Also provided is a food, a feed, or a processed product comprising theplant part of tomato variety NUN 00316 TOP, wherein the plant part is atomato fruit or part thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the fruit of tomato variety NUN 00316 TOP.

FIG. 2 shows the fruit comparison of tomato variety NUN 00316 TOP andthe Reference Variety.

FIG. 3 shows the cross-section and locule comparison of tomato varietyNUN 00316 TOP and the Reference Variety.

FIG. 4 shows the stem scar comparison of tomato variety NUN 00316 TOPand the Reference Variety.

FIG. 5 shows the stem end shape of tomato variety NUN 00316 TOP and theReference Variety.

FIG. 6 shows the blossom scar comparison of tomato variety NUN 00316 TOPand the Reference Variety.

FIG. 7 shows the blossom end shape of tomato variety NUN 00316 TOP andthe Reference Variety.

FIG. 8 shows the fruit cluster comparison of tomato variety NUN 00316TOP and the Reference Variety.

FIG. 9 shows the leaf comparison of tomato variety NUN 00316 TOP and theReference Variety.

DEFINITIONS

“Tomato” refers herein to plants of the species Solanum lycopersicum, ora closely related species, and fruits thereof. Solanum lycopersicum isalso known as Lycopersicon lycopersicum (L.) H. Karst. or Lycopersiconesculentum Mill. The most commonly eaten part of a tomato is the fruitor berry.

“Cultivated tomato” refers to plants of Solanum lycopersicum, or aclosely related species (e.g., varieties, breeding lines or cultivars ofthe species S. lycopersicum as well as crossbreds thereof, or crossbredswith other Solanum species), cultivated by humans and having goodagronomic characteristics.

The terms “tomato plant designated NUN 00316 TOP,” “NUN 00316 TOP,” “NUN00316,” “NUN 00316 F1,” “00316 TOP,” “tomato 00316,” or “N 6480” areused interchangeably herein and refer to a tomato plant of variety NUN00316 TOP, representative seed of which has been deposited underAccession Number NCIMB ______.

“Plant” includes the whole plant or any parts or derivatives thereof,having the same genetic makeup as the plant from which it is obtained.

“Plant part” includes any part of a plant, such as a plant organ (e.g.,harvested or non-harvested fruits), a plant cell, a plant protoplast, aplant cell tissue culture or a tissue culture from which a whole plantcan be regenerated, a plant cell that is intact in a plant, a clone, amicropropagation, plant callus, a plant cell clump, a plant transplant,a vegetative propagation, a seedling, a fruit, a harvested fruit, a partof a fruit, a leaf, a part of a leaf, pollen, an ovule, an embryo, apetiole, a shoot or a part thereof, a stem or a part thereof, a root ora part thereof, a root tip, a cutting, a seed, a part of a seed, ahypocotyl, a cotyledon, a scion, a graft, a stock, a rootstock, apistil, an anther, and a flower or a part thereof. Seed can be mature orimmature. Pollen or ovules may be viable or non-viable. Also, anydevelopmental stage is included, such as seedlings, cuttings prior orafter rooting, mature plants or leaves. Alternatively, a plant part mayalso include a plant seed which comprises one or two sets of chromosomesderived from the parent plant, e.g., from tomato variety NUN 00316 TOP.An F2 progeny produced from self-pollination of tomato variety NUN 00316TOP will thus comprise two sets of chromosomes derived from tomatovariety NUN 00316 TOP, while an F2 progeny derived fromcross-fertilization of tomato variety NUN 00316 TOP will comprise onlyone set of chromosomes from tomato variety NUN 00316 TOP, and the otherset of chromosomes from the other parent.

A “seed of tomato variety NUN 00316 TOP” refers to a tomato seed whichcan be grown into a plant of variety NUN 00316 TOP, wherein arepresentative sample of viable seed of tomato variety NUN 00316 TOP hasbeen deposited under Accession Number NCIMB ______. A seed can be in anystage of maturity, for example, a mature, viable seed, or an immature,non-viable seed. A seed comprises an embryo and maternal tissues.

An “embryo of tomato variety NUN 00316 TOP” refers to an “F1 hybridembryo” as present in a seed of tomato variety NUN 00316 TOP, arepresentative sample of said seed of tomato variety NUN 00316 TOP hasbeen deposited under Accession Number NCIMB ______.

A “seed grown on tomato variety NUN 00316 TOP” refers to a seed grown ona mature plant of variety NUN 00316 TOP or inside a fruit of tomatovariety NUN 00316 TOP. The “seed grown on NUN 00316 TOP” containstissues and DNA of the maternal parent, tomato variety NUN 00316 TOP.

A “fruit of NUN 00316 TOP” refers to a fruit containing maternal tissuesof tomato variety NUN 00316 TOP as has been deposited under AccessionNumber NCIMB ______. The fruit comprises pericarp, septa, epidermis,columella, locular cavity, vascular bundles and optionally seed.Pericarp, septa, epidermis, columella, locular cavity, vascular bundles,and seed coat of the seed are maternal tissues, e.g., they aregenetically identical to the plant on which they grow. In one aspect,the fruit contains seed grown on tomato variety NUN 00316 TOP. Inanother aspect, the fruit does not contain seed, i.e., the fruit isparthenocarpic. The skilled person is familiar with methods for inducingparthenocarpy. Those methods comprise chemically or genetically inducingparthenocarpy. Compounds suitable for chemically inducing parthenocarpycomprise auxins, gibberellins and cytokinins. Methods for geneticallyinducing parthenocarpy comprise the methods described in U.S. Pat. No.9,125,353, US 2002/0010953, U.S. Pat. No. 6,060,648, EP 1057401 and EP1428425, which are herein incorporated by reference in their entireties.

An “essentially homogeneous population of tomato seed” is a populationof seeds where at least 97%, 98%, 99% or more of the total population ofseed are seeds of tomato variety NUN 00316 TOP.

An “essentially homogeneous population of tomato plants” is a populationof plants where at least 97%, 98%, 99% or more of the total populationof plants are plants of tomato variety NUN 00316 TOP.

The phrase “essentially free from other seed” refers to a population ofseed where less than 3%, 2%, 1% or less of the total population of seedis seed that is not a tomato seed or, in another aspect, less than 3%,2%, 1% or less of the total population of seed is seed that is not seedof tomato variety NUN 00316 TOP.

“Harvest maturity” is referred to as the stage at which a tomato fruitis ripe or ready for harvest or the optimal time to harvest the fruitfor the market, for processing or for consumption. In one aspect,harvest maturity is the stage which allows proper completion of thenormal ripening.

“Harvested plant material” refers herein to plant parts (e.g., singlefruits or clusters of fruits detached from the whole plant), which havebeen collected for further storage and/or further use.

“Yield” means the total weight of all tomato fruits harvested perhectare of a particular line or variety. It is understood that “yield”expressed as weight of all tomato fruits harvested per hectare can beobtained by multiplying the number of plants per hectare times the“yield per plant”.

“Marketable yield” means the total weight of all marketable tomatofruits, especially fruit which is not cracked, damaged or diseased,harvested per hectare of a particular line or variety. A “marketablefruit” is a fruit that has commercial value.

“Rootstock” or “stock” refers to the plant selected for its roots, inparticular for the resistance of the roots to diseases or stress (e.g.,heat, cold, salinity etc.). Normally, the quality of the fruit of theplant providing the rootstock is less important.

“Scion” refers to a part of the plant attached to the rootstock. Thisplant is selected for its stems, leaves, flowers, or fruits. The scioncontains the desired genes to be duplicated in future production by thestock/scion plant and may produce the desired tomato fruit.

“Stock/scion” or grafted plant refers to a tomato plant comprising arootstock from one plant grafted to a scion from another plant.

“USDA descriptors” are the plant variety descriptors for tomato (Solanumlycopersicum or Lycopersicon esculentum Mill.) as published by the USDepartment of Agriculture, Agricultural Marketing Service, Plant VarietyProtection Office, Beltsville (June 2015) and which can be downloadedfrom the world wide web at ams.usda.gov underservices/plant-variety-protection/pvpo-c-forms under tomato. “Non-USDAdescriptors” are other descriptors suitable for describing tomato.

“UPOV descriptors” are the plant variety descriptors described fortomato in the “Guidelines for the Conduct of Tests for Distinctness,Uniformity and Stability, TG/44/11 (Geneva 2011, revised Oct. 30, 2018),as published by UPOV (International Union for the Protection of NewVarieties and Plants, and which can be downloaded from the world wideweb at upov.int/ under edocs/tgdocs/en/tg044.pdf and is hereinincorporated by reference in its entirety. Likewise, “UPOV methods” todetermine specific parameters for the characterization of tomato aredescribed at upov.int.

“Calibration Manual: DUS Test for Tomato” refers to the calibration bookfor tomato which provides guidance for describing a tomato variety, aspublished by Naktuinbow (Netherlands) and NCSS/NARO (Japan), January2020 and based on the UPOV TG/44/11.

“RHS” refers to the Royal Horticultural Society of England whichpublishes an official botanical color chart quantitatively identifyingcolors according to a defined numbering system. The chart may bepurchased from Royal Horticulture Society Enterprise Ltd RHS Garden;Wisley, Woking; Surrey GU236QB, UK, e.g., the RHS color chart: 2007.

“Reference Variety” refers herein to variety NUN 00241 TOP, a commercialvariety from Nunhems B.V., with commercial name N 6428, which has beenplanted in a trial together with tomato variety NUN 00316 TOP. Thecharacteristics of tomato variety NUN 00316 TOP are compared with thecharacteristics of the Reference Variety as shown in Tables 2-3. Thedistinguishing characteristics between tomato variety NUN 00316 TOP andthe Reference Variety are shown in Table 4.

A plant having “all the physiological and morphological characteristics”of a referred-to-plant means a plant showing the physiological andmorphological characteristics of the referred-to-plant when grown underthe same environmental conditions, preferably in the same experiment;the referred-to-plant can be a plant from which it was derived, e.g.,the progenitor plant, the parent, the recurrent parent, the plant usedfor tissue- or cell culture, etc. A physiological or morphologicalcharacteristic can be a numerical characteristic or a non-numericalcharacteristic. In one aspect, a plant has “all but one, two, or threeof the physiological and morphological characteristics” of areferred-to-plant, or “all the physiological and morphologicalcharacteristics” of Tables 1-3 or “all or all but one, two or three ofthe physiological and morphological characteristics” of Tables 1-3.

The physiological and/or morphological characteristics mentioned aboveare commonly evaluated at significance levels of 1%, 5%, or 10% if theyare numerical (quantitative), or for having an identical degree (ortype) if not numerical (not quantitative), if measured under the sameenvironmental conditions. For example, a progeny plant or a Single LocusConverted plant or a mutated plant of variety NUN 00316 TOP may have oneor more (or all) of the essential physiological and/or morphologicalcharacteristics of said variety listed in Tables 1-3, as determined atthe 5% significance level (i.e., p<0.05) for numerical characteristicsand determined by type or degree for non-numerical characteristics, whengrown under the same environmental conditions.

“Distinguishing characteristics” or “distinguishing morphological and/orphysiological characteristics” refers herein to the characteristicswhich distinguish (i.e., are different) between the new variety andother tomato varieties, such as the Reference Variety, when grown underthe same environmental conditions. The distinguishing characteristicsbetween tomato variety NUN 00316 TOP are described in Table 4. Whencomparing tomato variety NUN 00316 TOP with different varieties, thedistinguishing characteristics may be different. In one aspect, thedistinguishing characteristics may therefore include at least one, two,three or more (or all) of the characteristics listed in Tables 1-3. Allnumerical distinguishing characteristics are statistically significantlydifferent at p<0.05 between tomato variety NUN 00316 TOP, and the othervariety. All non-numerical distinguishing characteristics are different(in type or degree) between tomato variety NUN 00316 TOP and the othervariety.

Tomato variety NUN 00316 TOP has the following distinguishingcharacteristics when compared to the Reference Variety as shown in Table4: 1) shallowly toothed or scalloped margin of leaflets; 2) strongermarginal leaf rolling or wiltiness; 3) shorter leaf; 4) narrower leaf;5) smaller petiole; 6) lighter green leaf color; 7) shorter pedicel fromjoint to calyx; 8) larger fruit diameter; 9) heavier fruit; 10) three tofour locules; 11) lighter green fruit base color at mature-green stage;12) lower % soluble solids; 13) lighter intensity of green colorexcluding shoulder before maturity; 14) lighter yellow green color ofgreen fruit; 15) larger fruit size; 16) obovate fruit shape inlongitudinal section; 17) stronger depression at peduncle end; 18) flatto pointed shape of blossom end; 19) larger core in cross-section inrelation to total diameter; 20) darker red color of mature fruit; 21)darker red color of flesh at maturity; 22) larger pedicel; 23) resistantto Leveillula taurica; and 24) resistant to Pseudomonas syringae pvtomato, when the numerical characteristics are determined at 5%significance level and determined by type or degree for non-numericalcharacteristics when grown under the same environmental conditions.

Thus, a tomato plant “comprising the distinguishing characteristics oftomato variety NUN 00316 TOP” (such as a progeny plant) refers herein toa plant which does not differ significantly from said variety in thedistinguishing characteristics above. Therefore, in one aspect, a plant(such as a progeny plant of variety NUN 00316 TOP) is provided whichdoes not differ significantly from tomato variety NUN 00316 TOP in thedistinguishing characteristics above.

Similarity and differences between two different plant lines orvarieties can be determined by comparing the number of morphologicaland/or physiological characteristics (e.g., the characteristics aslisted in Tables 1-3) that are the same (i.e., statistically notsignificantly different) or that are different (i.e., statisticallysignificantly different) between the two plant lines or varieties whengrown under the same environmental conditions. A numericalcharacteristic is considered to be “the same” when the value for anumeric characteristic is not significantly different at the 1% (p<0.01)or 5% (p<0.05) significance level, using the T-Test, a standard methodknown to the skilled person. A non-numerical characteristic isconsidered to be “the same” when the values have the same “degree” or“type” when scored using USDA and/or UPOV descriptors for plants grownunder the same environmental conditions.

In one aspect, a statistical analysis of quantitative characteristicsshowing the degree of significance may be provided. Statisticalsignificance is the likelihood that a relationship between two or morevariables is caused by something other than chance, i.e., that thedifferences in the means for quantitative characteristics of tomatovariety NUN 00316 TOP and the Reference Variety are significant or dueto chance. For the purpose of proving differences or distinction betweentomato variety NUN 00316 TOP and the Reference Variety, a p-value of 5%(0.05) or lower is considered statistically significant. This means thatthere is only a 5% probability that the observed result could havehappened just by chance or random variation.

The statistical analysis is drawn from a small sample of at least 15plants or plant parts of tomato variety NUN 00316 TOP and the ReferenceVariety. Statistical points or parameters such as mean, minimum, median,maximum, and standard deviation are collected from the sample data toanalyze where the average is, how varied the data set is, and whetherthe data is skewed. For the purpose of determining whether the result ofa data set is statistically significant, a T-test is used, a statisticaltool for proving significance of means of the two groups (e.g., tomatovariety NUN 00316 TOP and the Reference Variety) at 5% significancelevel (p-value of 5% or 0.05).

The term “traditional breeding techniques” encompasses herein crossing,selfing, selection, doubled haploid production, embryo rescue,protoplast fusion, marker assisted selection, mutation breeding, etc. asknown to the breeder (e.g., methods other than geneticmodification/transformation/transgenic methods), by which, for example,a genetically heritable trait can be transferred from one tomato line orvariety to another.

“Variety,” “cultivated tomato,” or “cultivar” means a plant groupingwithin a single botanical taxon of the lowest known rank.

A “plant line” is, for example, a breeding line which can be used todevelop one or more varieties. A breeding line is typically highlyhomozygous.

“Hybrid variety” or “F1 hybrid” refers to the seeds harvested fromcrossing two inbred (nearly homozygous) parental lines. For example, thefemale parent is pollinated with pollen of the male parent to producehybrid (F1) seeds on the female parent.

“Progeny” as used herein refers to a plant obtained from a plantdesignated tomato variety NUN 00316 TOP. A progeny may be obtained byregeneration of cell culture or tissue culture or parts of a plant ofsaid variety or selfing of a plant of said variety or by producing seedsof a plant of said variety. In a further aspect, progeny may alsoencompass plants obtained from crossing of at least one plant of saidvariety with another tomato plant of the same variety or another varietyor (breeding) line, or with wild tomato plants. A progeny may comprise amutation or a transgene. A “first generation progeny” or is the progenydirectly derived from, obtained from, obtainable from or derivable fromthe parent plant by, e.g., traditional breeding methods (selfing and/orcross-pollinating) or regeneration (optionally combined withtransformation and mutation). Thus, a plant of variety NUN 00316 TOP isthe male parent, the female parent or both of a first generation progenyof that variety. Progeny may have all the physiological andmorphological characteristics of variety NUN 00316 TOP when grown underthe same environmental conditions. Using common breeding methods such asbackcrossing or recurrent selection, mutation or transformation, one ormore specific characteristics may be introduced into said variety, toprovide a plant comprising all but 1, 2, or 3 or more of themorphological and physiological characteristics of tomato variety NUN00316 TOP (e.g., as listed in Tables 1-3).

“Tissue culture” or “cell culture” refers to a composition comprisingisolated cells of the same or a different type or a collection of suchcells organized into parts of a plant. Tissue culture of various tissuesof tomato and regeneration of plants therefrom is well known and widelypublished (see, e.g., Bhatia et al. (2004), Plant Cell, Tissue and OrganCulture 78: 1-21). Similarly, methods of preparing cell cultures areknown in the art.

“Vegetative propagation,” “vegetative reproduction,” or “clonalpropagation” are used interchangeably herein and mean a method of takinga part of a plant and allowing that plant part to form at least roots,and also refer to the plant or plantlet obtained by that method.Optionally, the vegetative propagation is grown into a mature plant. Theskilled person is aware of what plant parts are suitable for use in themethod.

“Regeneration” refers to the development of a plant from cell culture ortissue culture or vegetative propagation.

“Crossing” refers to the mating of two parent plants. The termencompasses “cross-pollination” and “selfing”.

“Selfing” refers to self-pollination of a plant, i.e., the transfer ofpollen from the anther to the stigma of the same plant.

“Cross-pollination” refers to the fertilization by the union of twogametes from different plants.

“Backcrossing” is a traditional breeding technique used to introduce atrait into a plant line or variety. The plant containing the trait iscalled the donor plant and the plant into which the trait is transferredis called the recurrent parent. An initial cross is made between thedonor parent and the recurrent parent to produce a progeny plant.Progeny plants which have the trait are then crossed to the recurrentparent. After several generations of backcrossing and/or selfing therecurrent parent comprises the trait of the donor. The plant generatedin this way may be referred to as a “single trait converted plant.” Thetechnique can also be used on a parental line of a hybrid.

The terms “gene converted” or “conversion plant” or “single locusconverted plant” in this context refer to tomato plants which aredeveloped by traditional breeding techniques e.g., backcrossing, or viagenetic engineering or through mutation breeding, wherein essentiallyall of the desired morphological and physiological characteristics ofthe parent variety or line are recovered, in addition to the one or moregenes transferred into the parent via e.g., backcrossing technique(optionally including reverse breeding or reverse synthesis of breedinglines). It is understood that only the addition of a furthercharacteristic (e.g., addition of gene conferring a furthercharacteristic, such as a disease resistance gene), but also thereplacement/modification of an existing characteristic by a differentcharacteristic is encompassed herein (e.g., mutant allele of a gene canmodify the phenotype of a characteristic).

Likewise, a “Single Locus Converted (Conversion) Plant” refers to plantsdeveloped by plant breeding techniques comprising or consisting ofmutation and/or by genetic transformation and/or by traditional breedingtechniques, such as backcrossing, wherein essentially all of the desiredmorphological and physiological characteristics of a tomato variety arerecovered in addition to the characteristics of the single locus havingbeen transferred into the variety via the backcrossing technique. Incase of a hybrid, the gene may be introduced in the male or femaleparental line.

“Transgene” refers to a genetic locus comprising a DNA sequence whichhas been introduced into the genome of a tomato plant by transformation.A plant comprising a transgene stably integrated into its genome isreferred to as “transgenic plant.”

“Locus” (plural loci) refers to the specific location of a gene or DNAsequence on a chromosome. A locus may confer a specific trait.

“Genotype” refers to the genetic composition of a cell or organism.

“Allele” refers to one or more alternative forms of a gene locus. All ofthese loci relate to one trait. Sometimes, different alleles can resultin different observable phenotypic traits. However, many variations atthe genetic level result in little or no observable variation.

As used herein, the terms “resistance” and “tolerance” are usedinterchangeably to describe plants that show no symptoms orsignificantly reduced symptoms to a specified biotic pest, pathogen,abiotic influence or environmental condition compared to a susceptibleplant. These terms are optionally also used to describe plants showingsome symptoms but that are still able to produce marketable product withan acceptable yield.

DETAILED DESCRIPTION OF THE VARIOUS ASPECTS OF THE DISCLOSURE

The disclosure relates to the plant of variety NUN 00316 TOP, wherein arepresentative sample of seeds of said variety has been deposited underthe Budapest Treaty, with Accession Number NCIMB ______. NUN 00316 TOPis a determinate tomato variety of the viscosity type for the processingmarket and is suitable for growing in the open field.

The disclosure also provides a tomato plant or part thereof having allof the physiological and morphological characteristics of tomato varietyNUN 00316 TOP when grown under the same environmental conditions.

In another aspect, the plant of tomato variety NUN 00316 TOP or aprogeny thereof comprises resistance to Verticillium dahliae, Fusariumoxysporum s.p. lycopersici Race 2 (ex 3), Meloidogyne incognita,Pseudomonas syringae pv tomato, Leveillula taurica, and Tomato SpottedWilt Virus (TSWV), measured according to UPOV standards described inTG/44/11.

In another aspect, the plant of tomato variety NUN 00316 TOP or aprogeny plant thereof, comprises all of the following morphologicaland/or physiological characteristics (i.e., average values ofdistinguishing characteristics, as indicated on the USDA Objectivedescription of variety—tomato (unless indicated otherwise)) and shown inTables 1-3, where the numerical characteristics are determined at the 5%significance level and the non-numerical characteristics are determinedby type or degree for plants grown under the same environmentalconditions. A part of this plant is also provided.

The disclosure further provides a tomato plant which does not differfrom the physiological and morphological characteristics of the plant ofvariety NUN 00316 TOP as determined at the 1%, 2%, 3%, 4%, or 5%significance level for numerical characteristics and determined by typeor degree for non-numerical characteristics when grown under the sameenvironmental conditions. In a particular aspect, the plants aremeasured in the same trial (e.g., the trial is conducted as recommendedby USDA or UPOV). The disclosure also comprises a part of said plant,preferably a fruit or a part thereof.

The disclosure further relates to tomato variety NUN 00316 TOP, whichwhen compared to its Reference Variety has the following distinguishingcharacteristics as shown in Table 4: 1) shallowly toothed or scallopedmargin of leaflets; 2) stronger marginal leaf rolling or wiltiness; 3)shorter leaf; 4) narrower leaf; 5) smaller petiole; 6) lighter greenleaf color; 7) shorter pedicel from joint to calyx; 8) larger fruitdiameter; 9) heavier fruit; 10) three to four locules; 11) lighter greenfruit base color at mature-green stage; 12) lower % soluble solids; 13)lighter intensity of green color excluding shoulder before maturity; 14)lighter yellow green color of green fruit; 15) larger fruit size; 16)obovate fruit shape in longitudinal section; 17) stronger depression atpeduncle end; 18) flat to pointed shape of blossom end; 19) larger corein cross-section in relation to total diameter; 20) darker red color ofmature fruit; 21) darker red color of flesh at maturity; 22) largerpedicel; 23) resistant to Leveillula taurica; and 24) resistant toPseudomonas syringae pv tomato, when the numerical characteristics aredetermined at 5% significance level and determined by type or degree fornon-numerical characteristics when grown under the same environmentalconditions. Also encompassed are parts of the plant.

The morphological and/or physiological differences between two differentindividual plants described herein (e.g., between tomato variety NUN00316 TOP and a progeny of tomato variety NUN 00316 TOP) or between aplant of variety NUN 00316 TOP or progeny of said variety, or a planthaving all, or all but 1, 2, or 3, of the physiological andmorphological characteristics of tomato variety NUN 00316 TOP, (or all,or all but 1, 2, or 3 of the characteristics as listed in Tables 1-3)and another known variety can easily be established by growing saidvariety next to each other or next to the other variety (in the samefield, under the same environmental conditions), preferably in severallocations which are suitable for tomato cultivation, and measuringmorphological and/or physiological characteristics of a representativenumber of plants (e.g., to calculate an average value and to determinethe variation range/uniformity within the variety). For example, trialscan be carried out in Acampo California, USA (N 38 degrees 07′261″/W 121degrees 18′807″, USA), whereby various characteristics, for example,maturity, days from seeding to harvest, plant habit, plant attitude,stem branching, leaf color, inflorescence, shape of calyx, fruit shape,number of locules, fruit pattern, fruit color, flesh color, fruit size,disease resistance, insect resistance, can be measured and directlycompared for species of tomato.

Thus, the disclosure comprises tomato plant having one, two, or threephysiological and/or morphological characteristics which are differentfrom those of the plant of variety NUN 00316 TOP and which otherwise hasall the physiological and morphological characteristics of the plant ofvariety NUN 00316 TOP, e.g., determined at the 5% significance level fornumerical characteristics and determined by type or degree fornon-numerical characteristics for plants grown under the sameenvironmental conditions. In another aspect, the differentcharacteristic is the result of a mutation (e.g., spontaneous mutationor a human induced mutation through e.g., targeted mutagenesis ortraditional mutagenesis such as chemically or radiation inducedmutagenesis), or it is the result of transformation.

The disclosure also relates to a seed of tomato variety NUN 00316 TOP,wherein a representative sample of said seed has been deposited underthe Budapest Treaty, with Accession Number NCIMB ______.

In another aspect, a seed of hybrid variety NUN 00316 TOP is obtainableby crossing the male parent of said variety with the female parent ofsaid variety and harvesting the seeds produced on the female parent. Theresultant seeds of said variety can be grown to produce plants of saidvariety.

In another aspect, the disclosure provides a tomato plant grown from aseed of tomato variety NUN 00316 TOP and a plant part thereof.

In another aspect, the disclosure provides for a tomato plant part ofvariety NUN 00316 TOP, preferably a fruit or part thereof, arepresentative sample of seed from said variety has been deposited underthe Budapest Treaty, with Accession Number NCIMB ______.

In another aspect, the different characteristic(s) is/are result ofbreeding with tomato variety NUN 00316 TOP and selection of progenyplant comprising 1, 2, or 3 characteristics which are different than intomato variety NUN 00316 TOP.

Also provided is a plant of tomato variety NUN 00316 TOP, or a fruit, orother plant part thereof, produced from a seed, wherein a representativesample of said seeds has been deposited under the Budapest Treaty, withAccession Number NCIMB ______.

Also provided is a plant part obtained from variety NUN 00316 TOP,wherein said plant part is a fruit, a harvested fruit, a part of afruit, a leaf, a part of a leaf, pollen, an ovule, a cell, a petiole, ashoot or a part thereof, a stem or a part thereof, a root or a partthereof, a root tip, a cutting, a seed, a part of a seed, seed coat oranother maternal tissue which is part of a seed grown on said variety, ahypocotyl, a cotyledon, a scion, a stock, a rootstock, a pistil, ananther, and a flower or a part thereof. Such plant parts may be suitablefor sexual reproduction (e.g. a pollen, a flower or part thereof),vegetative reproduction (e.g., a cutting, a root, a stem, a cell, aprotoplast, a leaf, a cotyledon, a hypocotyl, a cell, a root, a roottip, an anther, a flower, a seed, or a stem). Fruits are particularlyimportant plant parts. Fruits may be parthenocarpic, or seedless, orcontain immature and/or nonviable seeds.

In a further aspect, the plant part obtained from variety NUN 00316 TOPis a cell, optionally a cell in a cell or tissue culture. That cell maybe grown into a plant of variety NUN 00316 TOP. A part of tomato varietyNUN 00316 TOP (or of progeny of that variety or of a plant having allphysiological and/or morphological characteristics but one, two, orthree which are different from those of tomato variety NUN 00316 TOP)further encompasses any cells, tissues, or organs obtainable from theseedlings or plants in any stage of maturity.

The disclosure also provides a tissue or cell culture comprising cellsof tomato variety NUN 00316 TOP. Such a tissue culture can, for example,be grown on plates or in liquid culture, or be frozen for long termstorage. The cells of tomato variety NUN 00316 TOP used to start theculture can be selected from any plant part suitable for vegetativereproduction, or, in a particular aspect, can be one or more of anembryo, a meristem, a cotyledon, a hypocotyl, pollen, a leaf, an anther,a root, a root tip, a pistil, a petiole, a flower, a fruit, seed, or astem of tomato variety NUN 00316 TOP. In another particular aspect, thetissue culture does not contain somaclonal variation or has reducedsomaclonal variation. The skilled person is familiar with methods toreduce or prevent somaclonal variation, including regular reinitiation.

In one aspect, the disclosure provides a tomato plant regenerated fromthe tissue or cell culture of tomato variety NUN 00316 TOP, wherein theregenerated plant is not significantly different from tomato variety NUN00316 TOP in all, or all but one, two, or three, of the physiologicaland morphological characteristics, e.g., determined at the 5%significance level for numerical characteristics and determined by typeor degree for non-numerical characteristics when grown under the sameenvironmental conditions. Optionally, the plant has one, two, or threeof the physiological and morphological characteristics that are affectedby a mutation or transformation with a transgene.

In another aspect, the disclosure provides a tomato plant regeneratedfrom the tissue or cell culture of tomato variety NUN 00316 TOP, whereinthe plant has all of the physiological and morphological characteristicsof said variety, e.g., determined at the 5% significance level fornumerical characteristics and determined by type or degree fornon-numerical characteristics when grown under the same environmentalconditions. Similarity or difference of a characteristic is determinedby measuring the characteristics of a representative number of plantsgrown under the same environmental conditions, determining whethertype/degree characteristics are the same or different and determiningwhether numerical characteristics are different at the 5% significancelevel.

Tomato variety NUN 00316 TOP, or its progeny, or a plant having allphysiological and/or morphological characteristics but one, two, orthree which are different from those of tomato variety NUN 00316 TOP,can also be reproduced using vegetative reproduction methods. Therefore,the disclosure provides for a method of producing a plant, or plant partof variety NUN 00316 TOP, comprising vegetative propagation of tomatovariety NUN 00316 TOP. Vegetative propagation comprises regenerating awhole plant from a plant part of variety NUN 00316 TOP, from a progenyor from or a plant having all physiological and/or morphologicalcharacteristics of said variety but one, two, or three differentcharacteristics, such as a cutting, a cell culture, or a tissue culture.

The disclosure also provides methods of vegetatively propagating a partof the plant of tomato variety NUN 00316 TOP. In certain aspects, themethod comprises: (a) collecting tissue or cells capable of beingpropagated from tomato variety NUN 00316 TOP; (b) cultivating saidtissue or cells to obtain proliferated shoots; and (c) rooting saidproliferated shoots, to obtain rooted plantlets. Steps (b) and (c) mayalso be reversed, i.e., first cultivating said tissue to obtain rootsand then cultivating the tissue to obtain shoots, thereby obtainingrooted plantlets. The rooted plantlets may then be further grown, toobtain plants. In one aspect, the method further comprises step (d)growing plants from said rooted plantlets. Therefore, the method alsocomprises regenerating a whole plant from said part of tomato varietyNUN 00316 TOP. In a particular aspect, the part of the plant to bepropagated is is a cutting, a cell culture, or a tissue culture.

The disclosure also provides for a vegetatively propagated plant ofvariety NUN 00316 TOP (or from progeny of tomato variety NUN 00316 TOPor from or a plant having all but one, two, or three physiologicaland/or morphological characteristics of said variety), wherein the planthas all of the morphological and physiological characteristics of tomatovariety NUN 00316 TOP, e.g., determined at the 5% significance level fornumerical characteristics and determined by type or degree fornon-numerical characteristics for plants grown under the sameenvironmental conditions. In another aspect, the propagated plant hasall but one, two, or three of the morphological and physiologicalcharacteristics of tomato variety NUN 00316 TOP, e.g., determined at the5% significance level for numerical characteristics and determined bytype or degree for non-numerical characteristics for plants grown underthe same environmental conditions. A part of said propagated plant orsaid propagated plant with one, two, or three differences is alsoprovided. In another aspect, the propagated plant has all or all butone, two, or three of the morphological and physiologicalcharacteristics of tomato variety NUN 00316 TOP (e.g., as listed inTables 1-3).

In another aspect, the disclosure provides a method for producing atomato plant part, preferably a fruit, comprising growing the plant ofvariety NUN 00316 TOP until it sets at least one fruit, and collectingthe fruit. Preferably, the fruit is collected at harvest maturity.

In another aspect, the fruit is collected when the seed is ripe. A plantof variety NUN 00316 TOP can be produced by seeding directly in the soil(e.g., field) or by germinating the seeds in controlled environmentconditions (e.g., greenhouses, hydroponic cultures, etc.) and optionallythen transplanting the seedlings into the field. For example, the seedcan be sown into prepared seed beds where they will remain for theentire production the crop (see, e.g., Hartz, et. al., University ofCalifornia Division of Agriculture and Natural Resources, Publication7228, 1-5). Tomatoes can be grown with a support system such as poles(i.e., stakes) to keep the fruit from touching the ground or as busheswithout support. Alternatively, plastic row covers can also be used tocontrol the temperature. Mulches or plastic tunnels can also be used toprotect the plant from frost (see, e.g., Le Strange, et. al., Universityof California Division of Agriculture and Natural Resources, Publication8017, 1-8). Tomato can also be grown entirely in greenhouses or nethouses. Moreover, said variety can be grown in hydroponic cultures asdescribed herein in, e.g., US 2008/0222949, which is herein incorporatedby reference in its entirety, and the skilled person is familiar withvarious type of hydroponic cultures.

In another aspect, the plant and plant parts of tomato variety NUN 00316TOP and progeny of said variety, e.g., grown from seeds, produced bysexual or vegetative reproduction, regenerated from the above-describedplant parts, or regenerated from cell or tissue culture of the tomatovariety NUN 00316 TOP, in which the reproduced (seed propagated orvegetatively propagated) plant has all of the physiological andmorphological characteristics of tomato variety NUN 00316 TOP, e.g., aslisted in Tables 1-3. In one aspect, said progeny of tomato variety NUN00316 TOP can be modified in one, two, or three characteristics, inwhich the modification is a result of mutagenesis or transformation witha transgene.

In another aspect, the disclosure provides a progeny plant of varietyNUN 00316 TOP such as a progeny plant obtained by further breeding oftomato variety NUN 00316 TOP. Further breeding with tomato variety NUN00316 TOP includes selfing that variety one or more times and/orcross-pollinating tomato variety NUN 00316 TOP with another tomato plantor variety one or more times. In particular, the disclosure provides fora progeny plant that retains all the essential morphological andphysiological characteristics of tomato variety NUN 00316 TOP or, inanother aspect, a progeny plant that retains all, or all but one, two,or three, of the morphological and physiological characteristics oftomato variety NUN 00316 TOP, optionally all or all but one, two, orthree of the characteristics as listed in Tables 1-3 when grown underthe same environmental conditions, determined at the 5% significancelevel for numerical characteristics and determined by type or degree fornon-numerical characteristics. In a particular aspect, the progeny is afirst generation progeny, i.e., the ovule or the pollen (or both) usedin the crossing is an ovule or pollen of variety NUN 00316 TOP, wherethe pollen comes from an anther and the ovule comes from an ovary oftomato variety NUN 00316 TOP. In another aspect, the disclosure providesfor a vegetative reproduction of the variety and a plant having all, orall but 1, 2, or 3 of the physiological and morphologicalcharacteristics of tomato variety NUN 00316 TOP (e.g., as listed inTables 1-3).

In still another aspect, the disclosure provides a method of producing atomato plant, comprising crossing a plant of tomato variety NUN 00316TOP with a second tomato plant at least once, allowing seed to developand optionally harvesting said progeny seed. The skilled person canselect progeny from said crossing. Optionally, the progeny (grown fromthe progeny seed) is crossed twice, thrice, or four, five, six, or seventimes, and allowed to set seed. In one aspect, the first “crossing”further comprises planting seeds of a first and a second parent tomatoplant, often in proximity so that pollination will occur; for example,mediated by insect vectors. Alternatively, pollen can be transferredmanually. Where the plant is self-pollinated, pollination may occurwithout the need for direct human intervention other than plantcultivation. After pollination the plant can produce seed.

The disclosure also provides a method for collecting pollen of tomatovariety NUN 00316 TOP, comprising collecting pollen from a plant ofvariety NUN 00316 TOP. Alternatively, the method comprises growing aplant of variety NUN 00316 TOP until at least one flower contains pollenand collecting the pollen. In particular aspect, the pollen is collectedwhen it is mature or ripe. A suitable method for collecting pollencomprises collecting anthers or the part of the anther that containspollen, for example, by cutting the anther or the part of the antheroff. Pollen can be collected in a container. Optionally, collectedpollen can be used to pollinate a tomato flower.

In yet another aspect, the disclosure provides a method of producing atomato plant, comprising selfing a plant of variety NUN 00316 TOP one ormore times, and selecting a progeny plant from said selfing. In oneaspect, the progeny plant retains all or all but one, two, or three ofthe morphological and physiological characteristics of tomato varietyNUN 00316 TOP when grown under the same environmental conditions. In adifferent aspect, the progeny plant comprises all (or all but one, two,or three) of the physiological and morphological characteristics oftomato variety NUN 00316 TOP as listed in Tables 1-3.

The disclosure also provides a method for developing a tomato plant in atomato breeding program, using a tomato plant described herein, or itsparts as a source of plant breeding material. Suitable plant breedingtechniques are recurrent selection, backcrossing, pedigree breeding,mass selection, mutation breeding and/or genetic marker enhancedselection. In one aspect, the method comprises crossing tomato varietyNUN 00316 TOP or progeny of said variety, or a plant comprising all but1, 2, or 3 or more of the morphological and physiologicalcharacteristics of tomato variety NUN 00316 TOP (e.g., as listed inTables 1-3), with a different tomato plant, and wherein one or moreoffspring of the crossing are subject to one or more plant breedingtechniques: recurrent selection, backcrossing, pedigree breeding, massselection, mutation breeding and genetic marker enhanced selection (seee.g., Vidaysky and Czosnek, (1998) Phytopathology 88(9): 910-4). Forbreeding methods in general, see, e.g., Principles of Plant Genetics andBreeding, (2007) George Acquaah, Blackwell Publishing, ISBN-13:978-1-4051-3646-4.

In one aspect, pedigree selection is used as a breeding method fordeveloping a tomato variety. Pedigree selection is also known as the“Vilmorin System of Selecton,” see, e.g., Allard, John Wiley & Sons,Inc., 1999, pp. 64-67. In general, selection is first practiced among F2plants. In the next season, the most desirable F3 lines are firstidentified, then desirable F3 plants within each line are selected. Thefollowing season and in all subsequent generations of inbreeding, themost desirable families are identified first, then desirable lineswithin the selected families are chosen. A family refers to lines thatwere derived from plants selected from the same progeny from thepreceding generation.

Thus, progeny in connection with pedigree selection are either thegeneration (seeds) produced from the first cross (F1) or selfing (S1),or any further generation produced by crossing and/or selfing (F2, F3,etc.) and/or backcrossing (BC1, BC2, etc.) one or more selected plantsof the F1 and/or S1 and/or BC1 generation (or plants of any furthergeneration, e.g., F2) with another tomato plant (and/or with a wildrelative of tomato). Progeny may have all the physiological andmorphological characteristics of tomato variety NUN 00316 TOP when grownunder the same environmental conditions and/or progeny may have (beselected for having) one or more of the distinguishing characteristicsof tomato variety NUN 00316 TOP.

In yet a further aspect, the disclosure provides for a method ofproducing a new tomato plant. The method comprises crossing tomatovariety NUN 00316 TOP, or a plant comprising all but 1, 2, or 3 of themorphological and physiological characteristics of tomato variety NUN00316 TOP (e.g., as listed in Tables 1-3), or a progeny plant thereof,either as male or as female parent, with a second tomato plant (or awild relative of tomato) one or more times, and/or selfing a tomatoplant of variety NUN 00316 TOP, or a progeny plant thereof, one or moretimes, and selecting progeny from said crossing and/or selfing. Thesecond tomato plant may, for example, be a line or variety of thespecies Solanum Lycopersicon, S. chilense, S. habrochaites, S. penelli,S. peruvianum, S. pimpinellifolium or other Solanum species.

In a further aspect, tomato variety NUN 00316 TOP is used in crosseswith other, different, tomato varieties to produce first generation (F1)tomato hybrid seeds and plants with superior characteristics. In aparticular aspect, the disclosure provides a tomato seed and a plantproduced by crossing a first parent tomato plant with a second parenttomato plant, wherein at least one of the first or second parent tomatoplant is tomato variety NUN 00316 TOP. In another aspect, the tomatoseed and plant produced are the first filial generation (F1) tomato seedand plants produced by crossing the plant of tomato variety NUN 00316TOP with another tomato plant.

The morphological and physiological characteristics (and thedistinguishing characteristics) of tomato variety NUN 00316 TOP areprovided, for example, in Tables 1-3. Encompassed herein is also a plantobtainable from tomato variety NUN 00316 TOP (e.g., by selfing and/orcrossing and/or backcrossing with said variety and/or progeny of saidvariety) comprising all or all but one, two or three of thephysiological and morphological characteristics of tomato variety NUN00316 TOP listed in Tables 1-3 as determined at the 5% significancelevel for numerical characteristics and determined by type or degree fornon-numerical characteristics when grown under the same environmentalconditions and/or comprising one or more (or all; or all except one, twoor three) characteristics when grown under the same environmentalconditions. The morphological and/or physiological characteristics mayvary somewhat with variation in the environment (such as temperature,light intensity, day length, humidity, soil, fertilizer use), which iswhy a comparison under the same environmental conditions is preferred.Colors can best be measured using the Royal Horticultural Society (RHS)Chart.

In another aspect, the disclosure provides a method of producing a plantderived from a tomato variety NUN 00316 TOP, comprising crossing a plantof variety NUN 00316 TOP either as a male or female parent with a secondplant or selfing tomato variety NUN 00316 TOP or vegetative reproductionof tomato variety NUN 00316 TOP and collecting seeds from said crossingor selfing or regenerating a whole plant from the vegetable cell-ortissue culture. Also provided are seeds and/or plants obtained by thismethod. All plants produced using tomato variety NUN 00316 TOP as aparent are within the scope of the disclosure including plant partsderived from tomato variety NUN 00316 TOP.

In further aspects, the method comprises growing a progeny plant of asubsequent generation and crossing the progeny plant of a subsequentgeneration with itself or a second plant and repeating the steps foradditional 3-10 generations to produce a plant derived from tomatovariety NUN 00316 TOP. The plant derived from tomato variety NUN 00316TOP may be an inbred line and the aforementioned repeating crossingsteps may be defined as comprising sufficient inbreeding to produce theinbred line. By selecting plants having one or more desirable traits ofthe line as well as potentially other selected traits.

The disclosure provides for methods of producing plants which retain allthe morphological and physiological characteristics of a plant describedherein. The disclosure also provides for methods of producing a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of tomato variety NUN 00316 TOP (e.g., aslisted in Tables 1-3), but which are still genetically closely relatedto said variety. The relatedness can, for example, be determined byfingerprinting techniques (e.g., making use of isozyme markers and/ormolecular markers such as Single-nucleotide polymorphism (SNP) markers,amplified fragment length polymorphism (AFLP) markers, microsatellites,minisatellites, Random Amplified Polymorphic DNA (RAPD) markers,restriction fragment length polymorphism (RFLP) markers and others). Aplant is “closely related” to variety NUN 00316 TOP if its DNAfingerprint is at least 80%, 90%, 95%, or 98% identical to thefingerprint of that variety. In a particular aspect AFLP markers areused for DNA fingerprinting (see, e.g., Vos et al. 1995, Nucleic AcidResearch 23: 4407-4414). A closely related plant may have a Jaccard'sSimilarity index of at least about 0.8, preferably at least about 0.9,0.95, 0.98 or more (see, e.g., Sharifova, S., et. al., (2013), Journalof Hort. Research, 21(1):83-89; Ince et al., (2010), Biochem. Genet.48:83-95; Parvathaneni et al., (2011), J. Crop Sci. Biotech, 14 (1):39-43; Pisanu, et. al., (2004), Acta Hort. 660, 83-89). The disclosurealso provides a plant and a variety obtained or selected by applyingthese methods on tomato variety NUN 00316 TOP. Such a plant may beproduced by crossing and/or selfing, or alternatively, a plant maysimply be identified and selected amongst plants of said variety, orprogeny of said variety, e.g. by identifying a variant within tomatovariety NUN 00316 TOP or within progeny of said variety (e.g., producedby selfing) which variant differs from the variety described herein inone, two or three of the morphological and/or physiologicalcharacteristics (e.g., in one, two or three distinguishingcharacteristics), e.g., those listed in Tables 1-3. In one aspect, thedisclosure provides a tomato plant having a Jaccard's Similarity indexwith tomato variety NUN 00316 TOP of at least 0.8, e.g. at least 0.85,0.9, 0.95, 0.98 or even at least 0.99.

In some aspects, the disclosure provides a tomato plant comprisinggenomic DNA having at least 95%, 96%, 97%, 98% or 99% sequence identitycompared to the genomic DNA sequence of a plant of variety NUN 00316 TOPas deposited under Accession Number NCIMB ______. In some aspects, thetomato plant further comprises all or all but 1, 2, or 3 of themorphological and physiological characteristics of tomato variety NUN00316 TOP (e.g., as listed in Tables 1-3). In other aspects, the tomatoplant is a hybrid or other derived from a seed or plant of NUN 00316TOP. In other aspects, the tomato plant comprises the distinguishingcharacteristics of tomato variety NUN 00316 TOP.

For the purpose of this disclosure, the “sequence identity” ofnucleotide sequences, expressed as a percentage, refers to the number ofpositions in the two optimally aligned sequences which have identicalresidues (×100) divided by the number of positions compared. A gap,i.e., a position in the pairwise alignment where a residue is present inone sequence but not in the other, is regarded as a position withnon-identical residues. A pairwise global sequence alignment of twonucleotide sequences is found by aligning the two sequences over theentire length according to the Needleman and Wunsch global alignmentalgorithm described in Needleman and Wunsch, 1970, J. Mol. Biol.48(3):443-53. A full implementation of the Needleman-Wunsch globalalignment algorithm is found in the needle program in The EuropeanMolecular Biology Open Software Suite (see, e.g., EMBOSS, Rice et al.,Trends in Genetics June 2000, vol. 16, No. 6. pp. 276-277).

In another aspect, the plant of variety NUN 00316 TOP may also bemutated (by e.g., irradiation, chemical mutagenesis, heat treatment,etc.) and mutated seeds or plants may be selected in order to change oneor more characteristics of said variety. Methods such as TILLING(Targeting Induced Local Lesions in Genomes) may be applied to tomatopopulations in order to identify mutants.

Similarly, tomato variety NUN 00316 TOP may be transformed andregenerated, whereby one or more chimeric genes are introduced into thevariety or into a plant comprising all but 1, 2, 3, or more of themorphological and physiological characteristics (e.g., as listed inTables 1-3). Many useful traits can be introduced into tomato varietyNUN 00316 TOP by e.g., crossing a tomato variety NUN 00316 TOP with atransgenic tomato plant comprising a desired transgene, as well as bydirectly introducing a transgene into tomato variety NUN 00316 TOP bygenetic transformation techniques.

Any pest or disease resistance genes may be introduced into a plant ofvariety NUN 00316 TOP, progeny of said variety or into a plantcomprising all but 1, 2, or 3 or more of the morphological andphysiological characteristics of tomato variety NUN 00316 TOP (e.g., aslisted in Tables 1-3). Resistance to one or more of the followingdiseases or pests may be introduced into plants described herein:Colorado potato beetle, Southern root knot nematode, Spider mites,Sugarfly beet army worm, Tobacco flea beetle, Tomato hornworm, Tomatofruitworm, Whitefly, Bacterial canker, Bacterial soft rot, Bacterialspeck, Bacterial wilt (Pseudomonas syringae pv. Tomato), Bacterial,Anthracnose (Gloeosporium piperatum), Brown rot or corky root(Pyrenochaeta lycopersici), Alternaria, Fusarium wilt (F. oxysporumraces), Gray leaf spot (Stemphylium spp.), Late blight (Phytophthorainfestans races), and Leaf mold (Cladosporium fulvum races), Nematode(Meloidogyne spp.), Verticillium Wilt (Verticillium dahliae), Ralstoniasolanacearum (Rs), Leveillula Taurica (Lt), and/or Oidium neolycopersici(On). Other resistance genes, against pathogenic viruses (e.g., TomatoMosaic Virus (ToMV), Curly Top Virus, Tomato Mottle Virus, Potato YVirus, Blotchey Ripening, Tobacco Etch Virus, the various Tobacco MosaicVirus races, Concentric cracking, Tomato Spotted Wilt Virus (TSWV),Tomato Yellow Leaf Curl Virus (TYLCV), Gold Fleck, Tomato Torrado Virus(ToTV)), Tomato Marchitez Virus (ToMaRV), Tomato Apex Necrosis Virus(ToANV), Tomato Brown Rugose Fruit Virus (ToBRFV), fungi, bacteria,nematodes, insects or other pests may also be introduced.

Genetic transformation may, therefore, be used to insert a selectedtransgene into the tomato plants of the disclosure described herein ormay, alternatively, be used for the preparation of transgenic tomatoplants which can be used as a source of the transgene(s), which can beintroduced into tomato variety NUN 00316 TOP by e.g., backcrossing. Agenetic trait which has been engineered into the genome of a particulartomato plant may then be moved into the genome of another tomato plant(e.g., another variety) using traditional breeding techniques which arewell-known in the art. For example, backcrossing is commonly used tomove a transgene from a transformed tomato variety into an alreadydeveloped tomato variety and the resulting backcross conversion plantwill then comprise the transgene(s).

Any DNA sequences, whether from a different species or from the samespecies, which are inserted into the genome using transformation, arereferred to herein collectively as “transgenes.” A “transgene” alsoencompasses antisense, or sense and antisense sequences capable of genesilencing. Thus, the disclosure also relates to transgenic plants oftomato variety NUN 00316 TOP. In some aspects, a transgenic plant oftomato variety NUN 00316 TOP may contain at least one transgene butcould also contain at least 1, 2, 3, 4, or more transgenes.

Plant transformation involves the construction of an expression vectorwhich will function in plant cells. Such a vector comprises DNAcomprising a gene under control of, or operatively linked to aregulatory element active in plant cells (e.g., promoter). Theexpression vector may contain one or more such operably linkedgene/regulatory element combinations. The vector may be in the form of aplasmid and can be used alone or in combination with other plasmids toprovided transformed tomato plants using transformation methods toincorporate transgenes into the genetic material of the tomato plant(s).Transformation can be carried out using standard methods, such asAgrobacterium tumefaciens mediated transformation, electroporation,biolistics particle delivery system, or microprojectile bombardment,followed by selection of the transformed cells and regeneration intoplants.

Plants can also be genetically engineered, modified, or manipulated toexpress various phenotypes of horticultural interest. Through thetransformation of tomato, the expression of genes can be altered toenhance disease resistance, insect resistance, herbicide resistance,stress tolerance, horticultural quality, and other traits.Transformation can also be used to insert DNA sequences which control orhelp control male sterility or fertility restoration. DNA sequencesnative to tomato as well as non-native DNA sequences can be transformedinto tomato and used to alter levels of native or non-native proteins.Various promoters, targeting sequences, enhancing sequences, and otherDNA sequences can be inserted into the genome for the purpose ofaltering the expression of proteins. Reduction of the specific activityof specific genes (also known as gene silencing or gene suppression) isdesirable for several aspects of genetic engineering in plants.

Genome editing is another method recently developed to geneticallyengineer plants. Specific modification of chromosomal loci or targetedmutation can be done through sequence-specific nucleases (SSNs) byintroducing a targeted DNA double strand break in the locus to bealtered. Examples of SSNs that have been applied to plants are fingernucleases (ZFNs), transcription activator-like effector nucleases(TALENs), engineered homing endonucleases or meganucleases, andclustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9 (Cas9), see, e.g., Songstad, et.al., Critical Reviews in Plant Sciences, 2017, 36:1, 1-23.

Thus, the disclosure also provides a method of producing a tomato planthaving a desired trait comprising mutating the plant or plant part ofvariety NUN 00316 TOP and selecting a plant with the desired trait,wherein the mutated plant retains all or all but one, two, or three ofthe physiological and morphological characteristics of tomato varietyNUN 00316 TOP, optionally as described in Tables 1-3, and contains thedesired trait and wherein a representative sample of seed of variety NUN00316 TOP has been deposited under Accession Number NCIMB ______. In afurther aspect, the desired trait yield, storage properties, color,flavor, size, firmness, fruit quality, enhanced nutritional quality,post-harvest quality, male sterility, herbicide tolerance, insectresistance, pest resistance, disease resistance, environmental stresstolerance, modified carbohydrate metabolism, modified protein metabolismor ripening, or wherein the desired trait comprises a mutation in any ofthe following genes acs2, acs4, rin, pp2c1, arf9, intense, myb12.

In one aspect, the disclosure provides a method for inducing mutation intomato variety NUN 00316 TOP comprising:

-   -   a. exposing the seed, plant, plant part, or cell of tomato        variety NUN 00316 TOP to a mutagenic compound or to radiation,        wherein a representative sample of seed of said tomato variety        has been deposited under Accession Number NCIMB ______;    -   b. selecting the seed, plant, plant part, or cell of tomato        variety NUN 00316 TOP, having a mutation; and    -   c. optionally growing and/or multiplying the seed, plant, plant        part, or cell of tomato variety NUN 00316 TOP, having the        mutation.

The disclosure also provides a method of producing a tomato plant havinga desired trait, wherein the method comprises transforming the tomatoplant with a transgene that confers the desired trait, wherein thetransformed plant contains the desired trait and otherwise retains allof the physiological and morphological characteristics of tomato varietyNUN 00316 TOP. Thus, a transgenic tomato plant is provided which isproduced by the method described above, wherein the plant otherwise hasall of the physiological and morphological characteristics of tomatovariety NUN 00316 TOP and the desired trait.

In another aspect, the disclosure provides a method of producing aprogeny of plant of variety NUN 00316 TOP further comprising a desiredtrait, said method comprising transforming the plant of tomato varietyNUN 00316 TOP with at least one transgene that confers the desired traitand/or crossing the plant of tomato variety NUN 00316 TOP with atransgenic tomato plant comprising a desired transgene so that thegenetic material of the progeny that resulted from the cross containsthe desired transgene(s). Also encompassed is the progeny produced bythis method.

A desired trait (e.g., gene(s) conferring pest or disease resistance, ortolerance for protection, etc.) can be introduced into tomato varietyNUN 00316 TOP, or progeny of said variety, by transforming said varietyor progeny of said variety with a transgene that confers the desiredtrait, wherein the transformed plant retains all or all but one, two orthree of the morphological and/or physiological characteristics ofvariety NUN 00316 TOP and contains the desired trait. In another aspect,the transformation or mutation confers a trait wherein the trait isyield, storage properties, color, flavor, size, firmness, fruit quality,enhanced nutritional quality, post-harvest quality, male sterility,herbicide tolerance, insect resistance, pest resistance, diseaseresistance, environmental stress tolerance, modified carbohydratemetabolism, modified protein metabolism or occurs in the intense gene.In a particular aspect, the specific transgene may be any known in theart or listed herein, including, a polynucleotide sequence conferringresistance to imidazolinone, sulfonylurea, glyphosate, glufosinate,triazine, benzonitrile, cyclohexanedione, phenoxy proprionic acid andL-phosphinothricin or a polynucleotide conferring resistance to Coloradopotato beetle, Southern root knot nematode, Spider mites, Sugarfly beetarmy worm, Tobacco flea beetle, Tomato hornworm, Tomato fruitworm,Whitefly, Bacterial canker, Bacterial soft rot, Bacterial speck,Bacterial wilt (Pseudomonas syringae pv. Tomato), Bacterial, Anthracnose(Gloeosporium piperatum), Brown rot or corky root (Pyrenochaetalycopersici), Alternaria, Fusarium wilt (F. oxysporum races), Gray leafspot (Stemphylium spp.), Late blight (Phytophthora infestans races), andLeaf mold (Cladosporium fulvum races), Nematode (Meloidogyne spp.),Verticillium Wilt (Verticillium dahliae), Ralstonia solanacearum (Rs),Leveillula Taurica (Lt), and/or Oidium neolycopersici (On). Otherresistance genes, against pathogenic viruses (e.g., Tomato Mosaic Virus(ToMV), Curly Top Virus, Tomato Mottle Virus, Potato Y Virus, BlotcheyRipening, Tobacco Etch Virus, the various Tobacco Mosaic Virus races,Concentric cracking, Tomato Spotted Wilt Virus (TSWV), Tomato YellowLeaf Curl Virus (TYLCV), Gold Fleck, Tomato Torrado Virus (ToTV)),Tomato Marchitez Virus (ToMaRV), Tomato Apex Necrosis Virus (ToANV),Tomato Brown Rugose Fruit Virus (ToBRFV), fungi, bacteria, nematodes,insects or other pests may also be introduced.

By crossing and/or selfing (one or more), single traits may beintroduced into tomato variety NUN 00316 TOP (e.g., using backcrossingbreeding schemes), while retaining the remaining morphological andphysiological characteristics of said variety and/or while retaining oneor more or all distinguishing characteristics. A single trait convertedplant may thereby be produced. For example, disease resistance genes maybe introduced, genes responsible for one or more quality traits, yield,etc. Both single genes (e.g., dominant or recessive) and one or moreQTLs (quantitative trait loci) may be transferred into tomato varietyNUN 00316 TOP by breeding with said variety.

Alternatively, a single trait converted plant or single locus convertedplant of variety NUN 00316 TOP may be produced by (i) geneticallytransforming or mutating cells of tomato variety NUN 00316 TOP; (ii)growing the cells into a plant; and (iii) optionally selecting a plantthat contains the desired single locus conversion. The skilled person isfamiliar with various techniques for genetically transforming a singlelocus in a plant cell, or mutating said cells.

In another aspect, the disclosure provides a method of introducing asingle locus conversion, single trait conversion, or a desired traitinto tomato variety NUN 00316 TOP, comprising introducing a single locusconversion, single trait conversion, or a desired trait in at least oneof the parents of tomato variety NUN 00316 TOP; and crossing theconverted parent with the other parent of tomato variety NUN 00316 TOPto obtain seed of tomato variety NUN 00316 TOP.

In another aspect, the step of introducing a single locus conversion,single trait conversion, or desired trait in at least one of the parentscomprises:

-   -   a. crossing the parental line of tomato variety NUN 00316 TOP        with a second tomato plant comprising the single locus        conversion, the single trait conversion or the desired trait;    -   b. selecting F1 progeny plants that contain the single locus        conversion, the single trait conversion or the desired trait;    -   c. crossing said selected progeny plants of step b) with the        parental line of step a), to produce a backcross progeny plant;    -   d. selecting backcross progeny plants comprising the single        locus conversion, the single trait conversion or the desired        trait and otherwise all or all but one, two or three of the        morphological and physiological characteristics the parental        line of step a) to produce selected backcross progeny plants;        and    -   e. optionally repeating steps c) and d) one or more times in        succession to produce selected second, third or fourth or higher        backcross progeny plants comprising the single locus conversion,        the single trait conversion or the desired trait and otherwise        all or all but one, two or three of the morphological and        physiological characteristics the parental line of step a) to        produce selected backcross progeny plants, when grown in the        same environmental conditions.

The disclosure further relates to plants obtained by this method.

In another aspect, the step of introducing a single locus conversion,single trait conversion, or a desired trait in at least one of theparents comprises:

-   -   a. obtaining a cell or tissue culture of cells of the parental        line of tomato variety NUN 00316 TOP;    -   b. genetically transforming or mutating said cells;    -   c. growing the cells into a plant; and    -   d. optionally selecting plants that contain the single locus        conversion, the single trait conversion or the desired trait.

In another aspect, the disclosure provides a method of introducing asingle locus conversion or single trait conversion or a desired traitinto tomato variety NUN 00316 TOP comprising:

-   -   a. obtaining a combination of a parental lines of tomato variety        NUN 00316 TOP, optionally through reverse synthesis of breeding        lines;    -   b. introducing a single locus conversion in at least one of the        parents of step a; and    -   c. crossing the converted parent with the other parent of step a        to obtain seed of tomato variety NUN 00316 TOP.

In another method, the step of introducing a single locus conversion,single trait conversion, or a desired trait in at least one of theparents comprises genetically transforming or mutating cells of theparental line of tomato variety NUN 00316 TOP; growing the cells into aplant; and optionally selecting plants that contain the single locusconversion, the single trait conversion or the desired trait.

In any of the above methods, where the single locus conversion concerns,single trait conversion, or a desired trait, the trait may be yield orpest resistance or disease resistance. In one aspect, the trait isdisease resistance and the resistance are conferred to Colorado potatobeetle, Southern root knot nematode, Spider mites, Sugarfly beet armyworm, Tobacco flea beetle, Tomato hornworm, Tomato fruitworm, Whitefly,Bacterial canker, Bacterial soft rot, Bacterial speck, Bacterial wilt(Pseudomonas syringae pv. Tomato), Bacterial, Anthracnose (Gloeosporiumpiperatum), Brown rot or corky root (Pyrenochaeta lycopersici),Alternaria, Fusarium wilt (F. oxysporum races), Gray leaf spot(Stemphylium spp.), Late blight (Phytophthora infestans races), and Leafmold (Cladosporium fulvum races), Nematode (Meloidogyne spp.),Verticillium Wilt (Verticillium dahliae), Ralstonia solanacearum (Rs),Leveillula Taurica (Lt), and/or Oidium neolycopersici (On). Otherresistance genes, against pathogenic viruses (e.g., Tomato Mosaic Virus(ToMV), Curly Top Virus, Tomato Mottle Virus, Potato Y Virus, BlotcheyRipening, Tobacco Etch Virus, the various Tobacco Mosaic Virus races,Concentric cracking, Tomato Spotted Wilt Virus (TSWV), Tomato YellowLeaf Curl Virus (TYLCV), Gold Fleck, Tomato Torrado Virus (ToTV)),Tomato Marchitez Virus (ToMaRV), Tomato Apex Necrosis Virus (ToANV),Tomato Brown Rugose Fruit Virus (ToBRFV), fungi, bacteria, nematodes,insects or other pests may also be introduced.

The disclosure also provides a plant having one, two, or threephysiological and/or morphological characteristics which are differentfrom those of tomato variety NUN 00316 TOP, and which otherwise has allthe physiological and morphological characteristics of said variety,wherein a representative sample of seed of said tomato variety has beendeposited under Accession Number NCIMB ______. In particular, variantswhich differ from tomato variety NUN 00316 TOP, in none, one, two, orthree of the characteristics mentioned in Tables 1-3 are encompassed.

The disclosure also provides a tomato plant comprising at least a firstset of the chromosomes of tomato variety NUN 00316 TOP, a sample of seedof said tomato variety has been deposited under Accession Number NCIMB______; optionally further comprising a single locus conversion or amutation, wherein said plant has essentially all of the morphologicaland physiological characteristics of the plant comprising at least afirst set of the chromosomes of said variety. In another aspect, thissingle locus conversion confers a trait, wherein the trait is yield,storage properties, color, flavor, size, firmness, fruit quality,enhanced nutritional quality, post-harvest quality, male sterility,herbicide tolerance, insect resistance, pest resistance, diseaseresistance, environmental stress tolerance, modified carbohydratemetabolism, modified protein metabolism or ripening, or wherein thedesired trait comprises a mutation in any of the following genes acs2,acs4, rin, pp2c1, arf9, intense, myb12.

In another aspect, the disclosure provides for a haploid plant and/or adoubled haploid plant of tomato variety NUN 00316 TOP, or of a planthaving all but one, two, or three physiological and/or morphologicalcharacteristics of tomato variety NUN 00316 TOP, or progeny of any ofthese. Haploid and doubled haploid (DH) plants can, for example, beproduced by cell or tissue culture and chromosome doubling agents andregeneration into a whole plant. For example, DH production chromosomedoubling may be induced using known methods, such as colchicinetreatment or the like. In one aspect, the method comprises inducing acell or tissue culture with a chromosome doubling agent and regeneratingthe cells or tissues into a whole plant.

In a further aspect, the disclosure comprises a method for makingdoubled haploid cells of tomato variety NUN 00316 TOP, comprising makingdoubled haploid cells from haploid cells from the plant or plant part oftomato variety NUN 00316 TOP with a chromosome doubling agent, such ascolchicine treatment (see, e.g., Nikolova V, Niemirowicz-Szczytt K(1996) Acta Soc Bot Pol 65:311-317).

The disclosure also provides for haploid plants and/or doubled haploidplants derived from tomato variety NUN 00316 TOP that, when combined,make a set of parents of tomato variety NUN 00316 TOP. The haploid plantand/or the doubled haploid plant of tomato variety NUN 00316 TOP can beused in a method for generating parental lines of tomato variety NUN00316 TOP.

The description also provides methods for determining the identity ofparental lines of plants described herein, in particular, the identityof the female line. US 2015/0126380, which is hereby incorporated byreference, relates to a non-destructive method for analyzing maternalDNA of a seed. In this method, the DNA is dislodged from the seed coatsurface and can be used to collect information on the genome of thematernal parent of the seed. This method for analyzing maternal DNA of aseed comprises contacting a seed with a fluid to dislodge DNA from theseed coat surface, and analyzing the DNA thus dislodged from the seedcoat surface using methods known in the art. The skilled person is thusable to determine whether a seed has grown on a plant of a plant ofvariety NUN 00316 TOP or is a progeny of said variety, because the seedcoat of the seed is a maternal tissue genetically identical to saidvariety. In one aspect, the present disclosure relates to a seed coatcomprising maternal tissue of tomato variety NUN 00316 TOP. In anotheraspect, the disclosure relates to a tomato seed comprising a maternaltissue of tomato variety NUN 00316 TOP. In another particular aspect,the disclosure provides for a method of identifying the female parentalline of tomato variety NUN 00316 TOP by analyzing the seed coat of aseed of that variety. In another aspect, the disclosure provides for amethod of determining whether a seed is grown on tomato variety NUN00316 TOP by analyzing the seed coat or another maternal tissue of saidseed.

In another aspect, a combination of a male and a female parental line oftomato variety NUN 00316 TOP can be generated by methods describedherein, for example, through reverse synthesis of breeding lines.

Using methods known in the art such as “reverse synthesis of breedinglines” or “reverse breeding”, it is possible to produce parental linesfor a hybrid plant such as tomato variety NUN 00316 TOP. A skilledperson can take any individual heterozygous plant (called a“phenotypically superior plant” inExample 2 of US 2015/0245570 herebyincorporated by reference in its entirety; NUN 00316 TOP is such plant)and generate a combination of parental lines (reverse breeding parentallines) that, when crossed, produce the variety NUN 00316 TOP. It is notnecessary that the reverse breeding parental lines are identical to theoriginal parental lines. Such new breeding methods are based on thesegregation of individual alleles in the spores produced by a desiredplant and/or in the progeny derived from the self-pollination of thatdesired plant, and on the subsequent identification of suitable progenyplants in one generation, or in a limited number of inbred cycles. Sucha method is known from US2015/0245570 or from Wijnker et al., NatureProtocols Volume: 9, Pages: 761-772 (2014) DOI:doi:10.1038/nprot.2014.049. Thus, the disclosure provides a method forproducing parental lines for a hybrid organism (e.g., tomato variety NUN00316 TOP), comprising in one aspect: a) defining a set of geneticmarkers present in a heterozygous form (H) in a partially heterozygousstarting organism; b) producing doubled haploid lines from spores of thestarting organism; c) genetically characterizing the doubled haploidlines thus obtained for the said set of genetic markers to determinewhether they are present in a first homozygous form (A) or in a secondhomozygous form (B); and d) selecting at least one pair of doubledhaploid lines that have complementary alleles for at least a subset ofthe genetic markers, wherein each member of the pair is suitable as aparental line for the hybrid organism.

In another aspect, the method for producing parental lines for hybridorganisms, e.g., of tomato variety NUN 00316 TOP, which when crossedreconstitute the genome of tomato variety NUN 00316 TOP, comprising:

-   -   a. defining a set genetic markers that are present a        heterozygous form (H) in a partially heterozygous starting        organism;    -   b. producing at least one further generation from the starting        organism by self-pollination (e.g., F2 or F3 generation);    -   c. selecting at least one pair of progeny organisms in which at        least one genetic marker from the set is present in a        complementary homozygous forms (B vs. A, or A vs. B); and    -   d. optionally repeating steps b) and c) until at least one pair        of progeny organisms that have complementary alleles for at        least a subset of the genetic markers has been selected as        parental lines for a hybrid.

Thus, in one aspect, the disclosure relates to a method of producing acombination of parental lines of a plant of tomato variety NUN 00316 TOPcomprising making doubled haploid cells from haploid cells from saidplant or a seed of that plant; and optionally crossing these parentallines to produce and collecting seeds. In another aspect, the disclosurerelates to a combination of parental lines produced by this method. Instill another aspect, the combination of parental lines can be used toproduce a seed or plant of variety NUN 00316 TOP when these parentallines are crossed. In still another aspect, the disclosure relates to acombination of parental lines from which a seed or a plant has all ofthe physiological and/or morphological characteristics of tomato varietyNUN 00316 TOP, e.g., when the numerical characteristics are determinedat the 5% significance level and determined by type or degree fornon-numerical characteristics for plants grown under the sameenvironmental conditions.

The disclosure also provides a method for producing parental lines forhybrid NUN 00316 TOP comprising: genetically characterizing a doubledhaploid line from tomato variety NUN 00316 TOP to determine whether oneor more genetic markers are present in a first homozygous form or in asecond homozygous form in said line, wherein the one or more geneticmarkers are present in a heterozygous form in tomato variety NUN 00316TOP; and selecting at least one pair of doubled haploid lines that havecomplementary alleles for the one or more the genetic markers, whereineach member of the pair is suitable as a parental line for a hybridorganism, optionally this method further comprises defining a set ofgenetic markers present in a heterozygous form in tomato variety NUN00316 TOP; and producing doubled haploid lines from tomato variety NUN00316 TOP. Doubled haploid lines generated as described herein can beused in such a method.

The disclosure also provides a combination of parental lines which, whencrossed, produce a seed or plant having all physiological and/ormorphological characteristics of tomato variety NUN 00316 TOP but one,two, or three which are different (when grown under the sameenvironmental conditions), as well as a seed or plant having allphysiological and/or morphological characteristics of tomato variety NUN00316 TOP, but one, two, or three which are different, e.g., when thenumerical characteristics are determined at the 5% significance leveland determined by type or degree for non-numerical characteristics forplants grown under the same conditions.

A part of tomato variety NUN 00316 TOP (or of progeny of said variety orof a plant having all physiological and/or morphological characteristicsbut one, two, or three which are different from those of said variety)encompasses any cells, tissues, organs obtainable from the seedlings orplants, such as but not limited to: a tomato fruit or a part thereof, acutting, hypocotyl, cotyledon, seed coat, pollen and the like. Suchparts can be stored and/or processed further. The disclosure furtherprovides for food or feed products comprising a part of tomato varietyNUN 00316 TOP or a part of progeny of said varieties, or a part of aplant having all but one, two or three physiological and/ormorphological characteristics of tomato variety NUN 00316 TOP,comprising one or more of such parts, optionally processed (such ascanned, chopped, cooked, roasted, in a sauce, in a sandwich, pasted,puréed or concentrated, juiced, frozen, dried, pickled, or powdered).

In another aspect, the disclosure provides a method of determining thegenotype of a plant described herein comprising detecting in the genome(e.g., a sample of nucleic acids) of the plant at least a firstpolymorphism or an allele. The skilled person is familiar with manysuitable methods of genotyping, detecting a polymorphism or detecting anallele including SNP (Single Nucleotide Polymorphism) genotyping,restriction fragment length polymorphism identification (RFLP) ofgenomic DNA, random amplified polymorphic detection (RAPD) of genomicDNA, amplified fragment length polymorphism detection (AFLP), polymerasechain reaction (PCR), DNA sequencing, allele specific oligonucleotide(ASO) probes, and hybridization to DNA microarrays or beads.Alternatively, the entire genome could be sequenced. The method may, incertain embodiments, comprise detecting a plurality of polymorphisms inthe genome of the plant, for example, by obtaining a sample of nucleicacid from a plant and detecting in said nucleic acids a plurality ofpolymorphisms. The method may further comprise storing the results ofthe step of detecting the plurality of polymorphisms on a computerreadable medium.

Also provided is a plant part obtainable from variety NUN 00316 TOP orfrom progeny of said variety or from a plant having all but one, two, orthree physiological and/or morphological characteristics which aredifferent from those of tomato variety NUN 00316 TOP, or from avegetatively propagated plant of variety NUN 00316 TOP (or from itsprogeny or from a plant having all or all but one, two, or threephysiological and/or morphological characteristics which are differentfrom those of tomato variety NUN 00316 TOP), wherein the plant part is afruit, a harvested fruit, a part of a fruit, a leaf, a part of a leaf,pollen, an ovule, a cell, a petiole, a shoot or a part thereof, a stemor a part thereof, a root or a part thereof, a root tip, a cutting, aseed, a part of a seed, seed-coat or another maternal tissue which ispart of a seed grown on tomato variety NUN 00316 TOP, or a hypocotyl, acotyledon, a scion, a stock, a rootstock, a pistil, an anther, or aflower or a part thereof.

A part of the plant of variety NUN 00316 TOP (or of progeny of saidvariety or of a plant having all physiological and morphologicalcharacteristics but one, two, or three which are different from those ofsaid variety) encompassed any cells, tissues, organs obtainable from theseedlings or plants, such as but not limited to a tomato fruit or partthereof, a cutting, a hypocotyl, a cotyledon, seed coat, or a pollen.

Such a plant part of tomato variety NUN 00316 TOP can be stored and/orprocessed further. The disclosure thus also provides for a food or afeed product comprising one or more of such parts from tomato varietyNUN 00316 TOP or from progeny of said variety, or from a derivedvariety, such as a plant having all or all but one, two, or three of thephysiological and morphological characteristics of tomato variety NUN00316 TOP. Preferably, the plant part is a tomato fruit or part thereofand/or an extract from the fruit of tomato variety NUN 00316 TOPcomprising at least a cell of tomato variety NUN 00316 TOP. The food orfeed product may be fresh or processed, e.g., dried, grinded, powdered,pickled, chopped, cooked, roasted, in a sauce, in a sandwich, pasted,puréed or concentrated, juiced, pickled, canned, steamed, boiled, fried,blanched and/or frozen, etc.

In another aspect, the disclosure provides for a tomato fruit of varietyNUN 00316 TOP, or a part of a fruit of said variety. The fruit can be inany stage of maturity, for example, immature or mature. In anotheraspect, the disclosure provides for a container comprising or consistingof a plurality of harvested tomato fruits or parts of fruits of saidvariety, or fruits of progeny thereof, or fruits of a derived variety.

Marketable tomato fruits are generally sorted by size and quality afterharvest. Alternatively, the tomato fruits can be sorted by expectedshelf life, pH or Brix.

In another aspect, the plant, plant part or seed of tomato variety NUN00316 TOP is inside a container, for example, containers such as cans,boxes, crates, bags, cartons, Modified Atmosphere Packaging, films(e.g., biodegradable films), etc. comprising a plant or a part of aplant (fresh and/or processed) or a seed of tomato variety NUN 00316TOP. In a particular aspect, the container comprises a plurality ofseeds of tomato variety NUN 00316 TOP, or a plurality of plant parts oftomato variety NUN 00316 TOP. The seed may be disinfected, primed and/ortreated with various compounds, such as seed coatings or crop protectioncompounds. The seed produces a plant of variety NUN 00316 TOP.

Tomato variety NUN 00316 TOP may also be grown for use as rootstocks(stocks) or scions. Typically, different types of tomatoes are graftedto enhance disease resistance, which is usually conferred by therootstock, while retaining the horticultural qualities usually conferredby the scion. It is not uncommon for grafting to occur betweencultivated tomato varieties and related tomato species. Methods ofgrafting and vegetative propagation are well-known in the art.

In another aspect, the disclosure provides to a plant comprising arootstock or scion of tomato variety NUN 00316 TOP.

All documents (e.g., patent publications) are herein incorporated byreference in their entirety, including the following cited references:

-   Naktuinbow (Netherlands) and NCSS/NARO (Japan), “Calibration Manual:    DUS Test for Tomato,” January 2020.-   UPOV, “Guidelines for the Conduct of Tests for Distinctness,    Uniformity and Stability”, TG/44/11 (Geneva 2011, revised Mar. 20,    2013), world-wide web at upov.int under edocs/tgdocs/en/tg044.pdf.-   US Department of Agriculture, Agricultural Marketing Service,    “Objective Description of Variety Tomato (Solanum lycopersicum or    Lycopersicon esculentum Mill)”, world wide web at    ams.usda.gov/services/plant-variety-protection/pypo-c-forms, under    tomato.-   Bhatia, P., et al., “Tissue Culture Studies of Tomato (Lycopersicum    esculentum)”, Plant Cell, Tissue and Organ Culture, 2004, vol. 78,    pp. 1-21.-   Hartz, T., et. al., “Processing Tomato Production in California,”    University of California Division of Agriculture and Natural    Resources, 1996, Publication 7228, pp. 1-5.-   Ince, A. G., et al., “Genetic Relationship Within and Between    Capsicum Species”, Biochem Genet, 2010, vol. 48, pp. 83-95.-   Le Strange, M., et. al., “Fresh-market Production in California,”    University of California Division of Agriculture and Natural    Resources, 2000, Publication 8017, pp. 1-8.-   Needleman, S. B., et. al., “A General Method Applicable to the    Search for Similarities in the Amino Acid Sequence of Two Proteins”,    Journal of Molecular Biology, 1970, vol. 48(3), pp. 443-53.-   Nikolova, V., et. al., “Diploidization of Cucumber (Cucumis sativus    L.) Haploids by Colchini Treatment”, Acta Societas Botanicorum    Poloniae, 1996, vol. 65, pp. 311-317.-   Pisanu, A. B., et. al., “Yield and Biometric Characteristics of 9    Clones Selected from the Population of “Spinoso sardo” Artichokes,    Acta Hort., 2004, ISHS 660, pp. 83-89.-   Rice, P., et al., “EMBOSS: The European Molecular Biology Open    Software Suite”, Trends in Genetics, 2000, vol. 16, Issue 6. pp.    276-277.-   Sharifova, S., et. al., “Assessment of Genetic Diversity in    Cultivated Tomato (Solanum lycopersicum L.) Genotypes Using RAPD    Primers”, Journal of Horticultural Research, 2013, vol. 21, no. 1,    pp. 83-89.-   Vidaysky, F., et. al., “Tomato Breeding Lines Resistant and Tolerant    to Tomato Yellow Leaf Curl Virus Issued from Lycopersicum hirsutum”,    The American Phytopathology Society, 1998, vol. 88, no. 9, pp.    910-914.-   Vos, P., et al., AFLP: A New Technique for DNA Fingerprinting 1995,    Nucleic Acids Research, 1995, vol. 23, No. 21, pp. 4407-4414.-   Wijnker, E., et al., Hybrid Recreation by Reverse breeding in    Arabidopsis thaliana, Nature Protocols, 2014, vol. 9, pp. 761-772.    DOI: doi: 10.1038/nprot.2014.049-   U.S. Pat. No. 9,125,353-   US 2002/0010953-   U.S. Pat. No. 6,060,648-   EP 1057401-   EP 1428425-   US 2008/0222949-   US 2015/0126380-   US 2015/0245570

Development of Tomato Variety NUN 00316 TOP

The hybrid NUN 00316 TOP was developed from a male and femaleproprietary inbred line of Nunhems based on yield and adaptability. Thefemale and male parents were crossed to produce hybrid (F1) seeds oftomato variety NUN 00316 TOP. The seeds of tomato variety NUN 00316 TOPcan be grown to produce hybrid plants and parts thereof (e.g., tomatofruit). The hybrid NUN 00316 TOP can be propagated by seeds orvegetatively.

The hybrid variety is uniform and genetically stable. This has beenestablished through evaluation of horticultural characteristics. Severalhybrid seed production events resulted in no observable deviation ingenetic stability. Coupled with the confirmation of genetic stability ofthe female and male parents the Applicant has concluded that tomatovariety NUN 00316 TOP is uniform and stable.

Deposit Information

A total of 625 seeds of the hybrid variety NUN 00316 TOP was made andaccepted according to the Budapest Treaty by Nunhems B.V. on ______ atthe NCIMB Ltd., Ferguson Building, Craibstone Estate, Bucksburn,Aberdeen AB21 9YA, United Kingdom (NCIMB). The deposit was assignedNCIMB Number ______. A statement indicating the viability of the samplehas been provided. A deposit of tomato variety NUN 00316 TOP and of themale and female parent line is also maintained at Nunhems B.V.

The deposit will be maintained in NCIMB for a period of 30 years, or 5years after the most recent request, or for the enforceable life of thepatent whichever is longer and will be replaced if it ever becomesnonviable during that period. Access to the deposits will be availableduring the pendency of this application to persons determined by theDirector of the U.S. Patent Office to be entitled thereto upon request.Subject to 37 C.F.R. § 1.808(b), all restrictions imposed by thedepositor on the availability to the public of the deposited materialwill be irrevocably removed upon the granting of the patent. Applicantdoes not waive any rights granted under this patent on this applicationor under the Plant Variety Protection Act (7 U.S.C. § 2321 et seq.).Accordingly, the requirements of 37 CFR § 1.801-1.809 have beensatisfied.

Characteristics of Tomato Variety NUN 00316 TOP

The most similar variety to tomato variety NUN 00316 TOP refers hereinto variety NUN 00241 TOP, a commercial variety from Nunhems B.V. withcommercial name N 6428.

In Table 1, the characteristics of tomato variety NUN 00316 TOP is shownbased on a trial in Acampo, Calif., USA in 2020 conducted under openfield conditions. For numerical characteristics averages, werecalculated. For non-numerical characteristics, the type/degree weredetermined.

In Tables 2-3, a comparison between tomato variety NUN 00316 TOP and theReference Variety is shown based on a trial in Acampo, Calif., USAconducted under open field conditions. Seeding date: Apr. 16, 2021;Transplanting date: May 21, 2021; Harvesting date: Aug. 9, 2021.

In Table 4, the distinguishing characteristics between tomato varietyNUN 00316 TOP and the Reference Variety are shown. For numericalcharacteristics averages, were calculated. For non-numericalcharacteristics, the type/degree were determined. For non-numericalcharacteristics, the type/degree were determined. Similarity anddifferences between two different plant lines or varieties can bedetermined by comparing the number of morphological and/or physiologicalcharacteristics (e.g., the characteristics as listed in Tables 1-3) thatare the same (i.e., statistically not significantly different) or thatare different (i.e., statistically significantly different) between thetwo plant lines or varieties when grown under the same environmentalconditions. A numerical characteristic is considered to be “the same”when the value for a numeric characteristic is not significantlydifferent at the 1% (p<0.01) or 5% (p<0.05) significance level, usingT-test, a standard method known to the skilled person. A non-numericalor “degree” or “type” characteristic is considered “the same” when thevalues have the same “degree” or “type” when scored using USDA and/orUPOV descriptors, if the plants are grown under the same environmentalconditions. In one aspect, a statistical analysis using T-test at 5%significance level is provided (see, Tables 5-15).

In another aspect, the disclosure provides a tomato plant having thephysiological and morphological characteristics of tomato variety NUN00316 TOP as presented in Tables 1-3 when grown under the sameenvironmental conditions.

TABLE 1 Characteristics of Tomato Variety NUN 00316 TOP, California.USA, 2020 Application Variety Characteristics (NUN 00316 TOP) Plant:Growth type: Determinate determinate, indeterminate, semi-determinant,semi indeterminant Inflorescence: Type: Simple Simple, Forked, CompoundType (2^(nd) and 3^(rd) truss): Mainly uniparous Peduncle: Abscissionlayer: Absent absent, present Fruit: Fruit shape: Plum or egg shapeflattened, slightly flattened, round, high round, pear, lengthened-pear,plum or egg shape, heart, lengthened cylindrical, angular Shape oflongitudinal section: Obovate flattened, oblate, circular, oblong,cylindrical, elliptic, cordate, ovate, obovate, pyriform, obcordateShape of blossom end: Tapered indented, flat, nippled, tapered Shape ofstem end: Flat flat, indented Shape of pistil scar: Dot dot, stellate,linear, irregular Ribbing at peduncle end: Absent or very weak absent orvery weak, weak, medium, strong, very strong Depression at peduncle end:Absent or very weak absent or very weak, weak, medium, strong, verystrong Fruit surface: Smooth smooth, slightly rough, moderately rough orribbed Glossiness of skin: Medium weak, medium, strong Color atmaturity: Red cream, yellow, orange, pink, red, brown, green Maturity:Time of maturity: Medium very early, early, medium, late, very lateResistances: Meloidoygne incognita (Mi) Resistant Verticillium dahliaeRace 0 Resistant Fusarium oxysporum f. sp. radicis lycopersici ResistantRace 2 (ex 3) Tomato Spotted Wilt Virus Resistant Leveillula tauricaResistant Pseudomonas syringae pv tomato Resistant

TABLE 2 Characteristics of Tomato Variety NUN 00316 TOP and theReference Variety (USDA Descriptors) based on California, USA Trial,2021 Application Variety Reference Variety Characteristics (NUN 00316TOP) (NUN 00241 TOP) Seedling: Anthocyanin in hypocotyl of 1-15 cmPresent Present seedling: absent, present Mature plant (at maximumvegetative development): Mature plant growth: Determinate Determinateindeterminate, determinate Form: Normal Normal lax, open; normal;compact; dwarf; brachytic Size of canopy: Medium Medium small, medium,large Mature plant habit: Semi-erect Semi-erect sprawling, semi-erect,erect Stem: Branching: Profuse Profuse sparse, intermediate, profuseBranching at cotyledonary or first leaf node: Present Present present,absent No. of nodes between early (1^(st)-2^(nd,) 2^(nd)-3^(rd)) Few Fewinflorescence: few (1-4), several (5-9), many (10 or more) No. of nodesbetween later developing Few Few inflorescences: few (1-4), several(5-9), many (10 or more) Leaf (mature leaf beneath the 3^(rd)inflorescence) Type of leaf blade: Bipinnate Bipinnate bipinnate,pinnate Margin of major leaflets: Shallowly toothed or Nearly entirenearly entire, shallowly toothed or scalloped scalloped, deeply toothedor cut toward base Marginal rolling or wiltiness: Moderate Slightabsent, slight, moderate, strong Surface of major leaflets: SmoothSmooth smooth, rugose (bumpy or veiny) Pubescence: Normal Normal Smooth(no long hairs), normal, hirsute, wooly Inflorescence (3^(rd)inflorescence) Type: Simple Simple simple, forked, compound Leafy or“running” inflorescence: Occasional Occasional absent, occasional,frequent Type (2^(nd) and 3^(rd) truss): Mainly uniparous Mainlyuniparous Flower: Corolla color: Yellow Yellow yellow, old gold, whiteor tan Fruit (3^(rd) fruit of 2^(nd) or 3^(rd) cluster): Fruit shape:Plum or egg shape Plum or egg shape flattened, slightly flattened,round, high round, pear, lengthened-pear, plum or egg shape, heart,lengthened cylindrical, angular Shape of transverse section: Round Roundround, flattened, angular, irregular Shape of blossom end: TaperedTapered indented, flat, nippled, tapered Shape of stem end: IndentedIndented flat, indented Shape of pistil scar: Dot Dot dot, stellate,linear, irregular Abscission layer: Absent Absent Present (pedicellate),absent (jointless) Point of detachment of fruit at At calyx detachmentAt calyx detachment harvest: at pedicel joint, at calyx attachmentLength of pedicel from joint to calyx 10.44 mm 13.89 mm attachment, mm:Diameter of fruit at widest point, mm: 54.85 mm 50.87 mm Mature fruitweight, g: 108.53 g 87.87 g Number of locules: Three to four Two tothree two to three, three and four, five or more Fruit surface: SmoothSmooth smooth, slightly rough, moderately rough or ribbed Fruit basecolor at mature-green stage: Yellow green Light green light green,yellow green, apple or medium green, light gray-green, dark green Fruitpattern at mature-green stage: Uniform green Uniform green uniformgreen, green-shouldered, radial stripes on sides of fruit Fruit color,full-ripe: Red Red white, yellow, orange, pink, red, brownish, greenish,red with purple, yellow with purple Flesh color, full-ripe: Red Redyellow, red/crimson, pink, orange, brownish, greenish Flesh color:Uniform Uniform uniform, with lighter and darker areas in walls Loculargel color of table-ripe fruit: Red Red green, yellow, red Stem scarsize: Medium Medium small, medium, large Fruit core: Present Presentcoreless, present Thickness of pericarp: Thick Thick thin, medium, thickThickness of pericarp, mm: 8.09 mm 7.90 mm Soluble solids as °Brix: 4.735.55 Adaptation: Culture: Field Field field, greenhouse, both Principleuse(s): Processing Processing home garden, fresh market, whole packcanning, concentrated products, other Resistances: Meloidoygne incognita(Mi) Resistant Resistant Verticillium dahliae Race 0 Resistant ResistantFusarium oxysporum f. sp. radicis Absent Absent lycopersici Race 0(ex 1) Fusarium oxysporum f. sp. radicis Absent Absent lycopersici Race1 (ex 2) Fusarium oxysporum f. sp. radicis Resistant Resistantlycopersici Race 2 (ex 3) Tomato Spotted Wilt Virus Resistant ResistantLeveillula taurica Resistant Absent Pseudomonas syringae pv tomatoResistant Absent

TABLE 3 Characteristics of Tomato Variety NUN 00316 TOP and theReference Variety (Non-USDA Descriptors) based on Acampo, California,USA Trial, 2021 Application Variety Reference Variety Characteristics(NUN 00316 TOP) (NUN 00241 TOP) Stem: Anthocyanin coloration of upperthird: Absent or very weak Absent or very weak absent or very weak,weak, medium, strong, very strong Leaf (mature leaf beneath the 3^(rd)inflorescence): Length: Short Medium very short, very short to short,short, short to medium, medium, medium to long, long, long to very long,very long Length, cm: 27.15 cm 33.46 cm Width: Medium Medium to broadvery narrow, very narrow to narrow, narrow, narrow to medium, medium,medium to broad, broad, broad to very broad, very broad Width, cm: 19.85cm 27.29 cm Petiole width, mm: 4.83 mm 6.72 mm Size of leaflets (inmiddle of leaf): Medium Medium very small, very small to small, small,small to medium, medium, medium to large, large to very large, verylarge Intensity of green color: Medium Dark very light, very light tolight, light, light to medium, medium, medium to dark, dark, dark tovery dark, very dark Glossiness (in middle third of plant): Very weakVery weak very weak, very weak to weak, weak, weak to medium, medium,medium to strong, strong, strong to very strong, very strong Blistering(in middle third of plant): Weak Weak very weak, very weak to weak,weak, weak to medium, medium, medium to strong, strong, strong to verystrong, very strong Size of blisters (in middle third of plant): MediumMedium very small, very small to small, small, small to medium, medium,medium to large, large to very large, very large Attitude of petioleleaflet in relation to main Semi-erect to horizontal Semi-erect tohorizontal axis (in middle third of plant): erect, erect to semi-erect,semi-erect, semi- erect to horizontal, horizontal, horizontal tosemi-drooping, semi-drooping, semi- drooping to drooping, drooping Leafstructure: Open Open open, intermediate, closed Color: Medium green Darkgreen light green, medium green, dark green RHS 137B RHS 137AAnthocyanin coloration of nerves: Absent Absent absent, present Flower:Color: Yellow Yellow yellow, orange Peduncle: Peduncle width, mm: 3.46mm 3.08 mm Fruit (3^(rd) fruit of 2^(nd) or 3^(rd) cluster): Greenshoulder (before maturity): Absent Absent absent, present Intensity ofgreen color excluding shoulder Very light Light (before maturity): verylight, very light to light, light, light to medium, medium, medium todark, dark, dark to very dark, very dark Green stripes (beforematurity): Absent Absent absent, present Green fruit color: Yellow greenYellow green RHS 145C RHS 144B Size: Medium to large Medium very small,very small to small, small to medium, medium, medium to large, large,large to very large, very large Ratio length/width: Moderately elongatedModerately elongated very compressed, moderately compressed, to veryelongated to very elongated medium, moderately elongated, very elongatedRatio length/width: 1.21 1.27 Shape in longitudinal section: ObovateElliptic flattened, oblate, circular, oblong, cylindrical, elliptic,cordate, ovate, obovate, pyriform, obcordate Ribbing at peduncle end:Absent or very weak Absent or very weak absent or very weak, very weakto weak, weak, weak to medium, medium, medium to strong, strong, strongto very strong, very strong Depression at peduncle end: Medium Weakabsent or very weak, very weak to weak, weak, weak to medium, medium,medium to strong, strong, strong to very strong, very strong Size ofpeduncle scar: Medium Medium very' small, very small to small, small tomedium, medium, medium to large, large, large to very large, very largeSize of blossom scar: Very small Very small very small, very small tosmall, small to medium, medium, medium to large, large, large to verylarge, very large Shape of blossom end: Flat to pointed Pointedindented, indented, flat, flat to pointed, conical Diameter of core incross-section in relation Medium Small to medium to total diameter: verysmall, very small to small, small to medium, medium, medium to large,large, large to very large, very large Cross-section: Round Round notround, round Color at maturity: Red Red cream, yellow, orange, pink,red, brown, RHS 175B RHS N172A green Color of flesh (at maturity): RedRed cream, yellow, orange, pink, red, brown, RHS 179A RHS 179B greenGlossiness of skin: Medium Medium weak, medium, strong Mature fruitlength, mm: 66.10 mm 64.75 mm Number of locules per fruit: 3.33 2.73Stem scar diameter, mm: 7.80 mm 7.60 mm

TABLE 4 Distinguishing Characteristics between Tomato Variety NUN 00316TOP and the Reference Variety Application Variety Reference VarietyCharacteristics (NUN 00316 TOP) (NUN 3351 TO) Leaf (mature leaf beneaththe 3^(rd) inflorescence): Margin of major leaflets: Shallowly toothedor Nearly entire nearly entire, shallowly toothed or scallopedscalloped, deeply toothed or cut toward base Marginal rolling orwiltiness: Moderate Slight absent, slight, moderate, strong Length:Short Medium very short, very short to short, short, short to medium,medium, medium to long, long, long to very long, very long Length, cm:27.15 cm 33.46 cm Width: Medium Medium to broad very narrow, very narrowto narrow, narrow, narrow to medium, medium, medium to broad, broad,broad to very broad, very broad Width, cm: 19.85 cm 27.29 cm Petiolewidth, mm: 4.83 mm 6.72 mm Intensity of green color: Medium Dark verylight, very light to light, light, light to medium, medium, medium todark, dark, dark to very dark, very dark Color: Medium green Dark greenlight green, medium green, dark green RHS 137B RHS 137A Fruit (3^(rd)fruit of 2^(nd) or 3^(rd) cluster): Length of pedicel from joint tocalyx 10.44 mm 13.89 mm attachment, mm: Diameter of fruit at widestpoint, mm: 54.85 mm 50.87 mm Mature fruit weight, g: 108.53 g 87.87 gNumber of locules: Three to four Two to three two to three, three andfour, five or more Fruit base color at mature-green stage: Yellow greenLight green light green, yellow green, apple or medium green, lightgray-green, dark green Soluble solids as °Brix: 4.73 5.55 Intensity ofgreen color excluding shoulder Very light Light (before maturity): verylight, very light to light, light, light to medium, medium, medium todark, dark, dark to very dark, very dark Green fruit color: Yellow greenYellow green RHS 145C RHS 144B Size: Medium to large Medium very small,very small to small, small to medium, medium, medium to large, large,large to very large, very large Shape in longitudinal section: ObovateElliptic flattened, oblate, circular, oblong, cylindrical, elliptic,cordate, ovate, obovate, pyriform, obcordate Depression at peduncle end:Medium Weak absent or very weak, very weak to weak, weak, weak tomedium, medium, medium to strong, strong, strong to very strong, verystrong Shape of blossom end: Flat to pointed Pointed indented, indented,flat, flat to pointed, conical Diameter of core in cross-section inrelation Medium Small to medium to total diameter: very small, verysmall to small, small to medium, medium, medium to large, large, largeto very large, very large Color at maturity: Red Red cream, yellow,orange, pink, red, brown, RHS 175B RHS N172A green Color of flesh (atmaturity): Red Red cream, yellow, orange, pink, red, brown, RHS 179A RHS179B green Pedicel diameter, mm: 3.46 mm 3.08 mm Disease Resistances:Leveillula taurica Resistant Absent Pseudomonas syringae pv tomatoResistant Absent

The results of the T-test show significant differences at 5%significance level between tomato variety NUN 00316 TOP and theReference Variety for leaf length, leaf width, petiole width, pedicellength, pedicel width, fruit diameter, fruit weight, and % solublesolids as shown in Tables 5-12.

Table 5 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p<0.001) on leaflength (cm) based on the results of the trial conducted in the US duringthe trial season 2021.

TABLE 5 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 22.80 29.30Max. 30.0 37.10 Median 27.80 33.30 Mean 27.15 33.46 Standard deviation2.16 1.91

Table 6 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p<0.001) on leafwidth (cm) based on the results of the trial conducted in the US duringthe trial season 2021.

TABLE 6 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 15.70 22.70Max. 23.10 31.40 Median 19.50 27.30 Mean 19.85 27.29 Standard deviation2.49 2.80

Table 7 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p<0.001) onpetiole width (mm) based on the results of the trial conducted in the USduring the trial season 2021.

TABLE 7 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 4.15 5.91 Max.5.94 7.83 Median 4.63 6.74 Mean 4.83 6.72 Standard deviation 0.55 0.53

Table 8 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p<0.001) onpedicel length (mm) based on the results of the trial conducted in theUS during the trial season 2021.

TABLE 8 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 8.78 12.57Max. 11.97 15.34 Median 10.65 13.68 Mean 10.44 13.89 Standard deviation0.94 0.86

Table 9 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p=0.014) onpedicel width (mm) based on the results of the trial conducted in the USduring the trial season 2021.

TABLE 9 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 2.95 2.40 Max.4.37 3.62 Median 3.37 3.15 Mean 3.46 3.08 Standard deviation 0.44 0.36

Table 10 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p<0.001) onfruit weight (g) based on the results of the trial conducted in the USduring the trial season 2021.

TABLE 10 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 86.0 66.0 Max.136.0 106.0 Median 108.0 90.0 Mean 108.53 87.87 Standard deviation 13.4112.13

Table 11 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p=0.004) onfruit diameter (mm) based on the results of the trial conducted in theUS during the trial season 2021.

TABLE 11 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 47.62 45.82Max. 62.20 55.47 Median 55.46 50.36 Mean 54.85 50.87 Standard deviation4.07 2.76

Table 12 shows a significant difference at 5% significance level betweentomato variety NUN 00316 TOP and the Reference Variety (p<0.001) on %soluble solids (°Brix) based on the results of the trial conducted inthe US during the trial season 2021.

TABLE 12 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 3.90 4.80 Max.5.50 6.30 Median 4.70 5.50 Mean 4.73 5.55 Standard deviation 0.50 0.40

The results of the T-test show no significant differences at 5%significance level between tomato variety NUN 00316 TOP and theReference Variety for fruit length, thickness of pericarp, and stem scardiameter as shown in Table 13-15.

Table 13 shows no significant difference at 5% significance levelbetween tomato variety NUN 00316 TOP and the Reference Variety (p=0.398)on fruit length (mm) based on the results of the trial conducted in theUS during the trial season 2021.

TABLE 13 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 60.34 54.24Max. 71.07 70.68 Median 66.99 66.17 Mean 66.10 64.75 Standard deviation3.38 5.06

Table 14 shows no significant difference at 5% significance levelbetween tomato variety NUN 00316 TOP and the Reference Variety (p=0.524)on thickness of pericarp (mm) based on the results of the trialconducted in the US during the trial season 2021.

TABLE 14 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 6.32 7.22 Max.9.91 8.94 Median 7.98 7.83 Mean 8.09 7.90 Standard deviation 1.03 0.45

Table 15 shows no significant difference at 5% significance levelbetween tomato variety NUN 00316 TOP and the Reference Variety (p=0.524)on stem scar diameter (mm) based on the results of the trial conductedin the US during the trial season 2021.

TABLE 15 Application Variety Reference Variety Statistical Parameters(NUN 00316 TOP) (NUN 00241 TOP) No. of samples 15 15 Min. 6.78 6.42 Max.9.31 9.81 Median 7.73 7.27 Mean 7.80 7.60 Standard deviation 0.63 1.0

1. A plant, plant part, or seed of tomato variety NUN 00316 TOP, wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 2. The plant part of claim 1, wherein said plant part is a fruit, a leaf, pollen, an ovule, a cell, a scion, a root, a rootstock, a cutting, or a flower.
 3. A seed that produces the plant of claim
 1. 4. A plant produced by growing the seed of claim
 1. 5. A tomato plant or part thereof having all the physiological and morphological characteristics of the plant of claim 1 when grown under the same environmental conditions.
 6. A tissue or cell culture of regenerable cells of the plant or plant part of claim
 1. 7. The tissue or cell culture according to claim 6, comprising cells or protoplasts obtained from a plant part suitable for vegetative reproduction, wherein the plant part is a meristem, a cotyledon, a hypocotyl, a pollen, a leaf, an anther, a root, a root tip, a pistil, a petiole, a flower, a fruit, a stem, or a stalk.
 8. A tomato plant regenerated from the tissue or cell culture of claim 6, wherein the plant has all of the physiological and morphological characteristics of the plant of tomato variety NUN 00316 TOP, when grown under the same environmental conditions, and wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 9. A method of producing the plant of claim 1 or a part thereof, said method comprising vegetative propagation of at least a part of the plant of tomato variety NUN 00316 TOP, wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 10. The method of claim 9, wherein said vegetative propagation comprises regenerating a whole plant from the plant part of tomato variety NUN 00316 TOP, wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 11. The method of claim 9, wherein said part is a cutting, a cell culture, or a tissue culture.
 12. A vegetatively propagated plant, or a part thereof, produced by the method of claim 9, wherein the plant or a part thereof has all of the physiological and morphological characteristics of the plant of tomato variety NUN 00316 TOP when grown under the same environmental conditions, and wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 13. A method of producing a tomato plant, said method comprising crossing the plant of claim 1 with a second tomato plant at least once, producing a progeny tomato plant from said crossing, and optionally allowing the progeny tomato plant to form seed.
 14. A method of producing a tomato plant, said method comprising selfing the plant of claim 1 one or more times, and selecting a progeny tomato plant from said selfing and optionally allowing the progeny tomato plant to form seed.
 15. A method of making doubled haploids of the plant of claim 1, said method comprising making double haploid cells from haploid cells of the plant or plant part of tomato variety NUN 00316 TOP, wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 16. A plant comprising the scion or rootstock of claim
 2. 17. A container comprising the plant, plant part, or seed of claim
 1. 18. A food, a feed, or a processed product comprising the plant part of claim
 2. 19. A method of introducing a desired trait into the plant of claim 1, said method comprises transforming the plant of claim 1 with a transgene that confers the desired trait, wherein the desired trait is male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, environmental stress tolerance, modified carbohydrate metabolism, or modified protein metabolism.
 20. A tomato plant produced by the method of claim 19, wherein the transformed plant otherwise comprises the desired trait and otherwise has all the physiological and morphological characteristics of tomato variety NUN 00316 TOP.
 21. A method of producing a tomato fruit, said method comprising growing the plant of claim 1 until it sets at least one fruit, and collecting the fruit.
 22. A fruit produced by the method of claim
 21. 23. A container comprising the tomato fruit collected by the method of claim
 21. 24. A method of producing a modified tomato plant having a desired trait, said method comprises mutating a plant or plant part of tomato variety NUN 00316 TOP and selecting the plant with a desired trait, wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 25. A tomato plant produced by the method of claim 24, wherein the modified plant comprises the desired trait and otherwise all the physiological and morphological characteristics of tomato variety NUN 00316 TOP and wherein the desired trait is male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, environmental stress tolerance, modified carbohydrate metabolism, or modified protein metabolism.
 26. A method of producing a tomato seed, said method comprises crossing tomato plants and harvesting the resultant seed, wherein at least one tomato plant in the cross is the plant of claim 1, wherein a representative sample of seed of said tomato variety NUN 00316 TOP has been deposited under Accession Number NCIMB ______.
 27. A method of introducing a single locus conversion into the plant of claim 1, comprising: a. crossing the plant of claim 1 with a second tomato plant comprising a desired single locus conversion to produce F1 progeny plants; b. selecting the F1 progeny plants that have the single locus conversion to produce selected F1 progeny plants; c. crossing selected F1 progeny plants with tomato variety NUN 00316 TOP to produce backcross progeny plants; d. selecting backcross progeny plants that have the single locus conversion and otherwise comprise all of the physiological and morphological characteristics of tomato variety NUN 00316 TOP to produce selected backcross progeny plants; and e. repeating steps (c) and (d) one or more times in succession to produce selected second or higher backcross progeny plants that comprise the single locus and otherwise comprise all of the physiological and morphological characteristics of tomato variety NUN 00316 TOP, wherein a representative sample of seed of said tomato variety has been deposited under Accession Number NCIMB ______.
 28. The method of claim 27, wherein the single locus conversion confers male sterility, herbicide tolerance, insect resistance, pest resistance, disease resistance, environmental stress tolerance, modified carbohydrate metabolism, or modified protein metabolism.
 29. A tomato plant produced by the method of claim 27, wherein the plant comprises the single locus conversion and otherwise has all the physiological and morphological characteristics of tomato variety NUN 00316 TOP.
 30. A method of producing a tomato plant derived from the plant of claim 1, comprising: a. preparing a progeny tomato plant from tomato variety NUN 00316 TOP by crossing the plant of claim 1 with itself or with a second tomato plant; b. crossing the progeny plant with itself or a second tomato plant to produce a seed of a progeny plant of the subsequent generation; c. growing a progeny plant of the subsequent generation from said seed and crossing the progeny plant of the subsequent generation with itself or a second tomato plant; and d. repeating steps (b) and (c) for a least one more generation to produce a tomato plant derived from tomato variety NUN 00316 TOP.
 31. A method of determining the genotype of the plant of claim 1, said method comprises obtaining a sample of nucleic acids from said plant, detecting in said nucleic acids a plurality of polymorphisms, thereby determining the genotype of the plant, and storing the results of detecting the plurality of polymorphisms on a computer readable medium.
 32. A method of developing a tomato plant in a tomato breeding program, said method comprising applying plant breeding techniques comprising recurrent selection, backcrossing, pedigree breeding, mass selection, mutation breeding, genetic marker enhanced selection, or genetic transformation to the plant of claim 1 or part thereof.
 33. A progeny of the tomato plant of claim 1, obtained by crossing the plant of tomato variety NUN 00316 TOP with itself or another tomato plant. 